FXR modulators

ABSTRACT

The present invention provides compounds, pharmaceutical compositions and methods that are useful in modulating the farnesoid X receptor (FXR). As FXR is involved in negatively controlling the expression level of cholesterol 7α-hydroxylase (cyp 7 a), the rate-limiting enzyme involved in the oxidative metabolism of cholesterol into bile acids, the compounds described herein find utility in treating diseases associated with abnormally high or low cholesterol levels. In certain aspects, the FXR modulators (e.g., antagonists) described herein block the negative feed-back downregulation of cyp 7 a expression produced by certain cholic acids, the endogenous ligands for FXR. Moreover, as FXR forms heterodimers with the retinoid X receptor (RXR) in some cell types, modulation of the level of FXR activity in cells has a wide range of effects on a variety of biological processes which are mediated by RXR or other RXR-interacting proteins such as PPARγ and PPARα. Thus, compounds described herein are useful in treating other biological activities such as obesity, diabetes, lipid associated disorders, cancer, inflammatory disorders, disorders involving a disrupted or dysfunctional epidermal barrier, and various other metabolic disorders.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims priority to U.S. Application No.60/230,585, filed Sep. 5, 2000, and 60/258,092, filed Dec. 22, 2000, theteachings of which are both incorporated herein by reference.

BACKGROUND OF THE INVENTION

[0002] The farnesoid X receptor (FXR), the peroxisomeproliferator-activated receptor α (PPARα), and the liver X receptor α(LXRα) are members of a superfamily of approximately 150 proteins thatbind to cis-acting elements in the promoters of their target genes andmodulate gene expression in response to hormone activators or ligands.For many of these receptors, the activators are known, while for others,termed “orphan receptors,” the activators are unknown. Furthermore, someof these receptors bind to their target genes as dimers consisting oftwo molecules of the same receptor (homodimers), while others bind asdimers consisting of one molecule each of two different receptors(heterodimers). Prominent among the latter are nuclear receptors thatrequire heterodimerization with the retinoid X receptor (RXR), asdisclosed in Yu et al. Cell 67:1251-1266 (1991). Members of this groupinclude the vitamin D receptor, the thyroid hormone receptor (T₃R), theretinoic acid receptor (RAR), FXR, the peroxisome proliferator-activatedreceptors (PPARs) and LXR α.

[0003] FXR was first reported by Forman and coworkers, Forman Cell81:687-693 (1995). This receptor is a protein having a relativemolecular mass (Mr) of approximately 54,000, and is a vertebratetranscription factor regulated by intracellular metabolites. Thereceptor is activated by certain farnesoids, i.e., farnesol, compoundsderived from farnesol, and/or compounds similar in structure tofarnesol. These farnesoids include farnesol, farnesal, farnesyl acetate,farnesoic acid, geranylgeraniol and juvenile hormone III.

[0004] FXR is a nuclear receptor thought to be involved in negativelycontrolling the expression level of cholesterol 7α-hydroxylase (cyp7a),the rate-limiting enzyme involved in the oxidative metabolism ofcholesterol into bile acids. As such, modulators of FXR activity willfind utility in diseases associated with abnormally high or lowcholesterol levels. Of particular value will be FXR antagonists, whichblock the negative feedback downregulation of cyp7a expression producedby certain cholic acids, the endogenous ligands for FXR. FXR is alsoinvolved in controlling the synthesis of isoprenoid derivatives(including cholesterol), and the proliferation of certain types ofcancerous cells, such as those derived from colon carcinomas.Additionally, since FXR forms heterodimers with RXR in some cell types,modulation of the level of FXR activity in a cell has a wide range ofeffects on a variety of biological processes which are mediated by RXRor other RXR-interacting proteins such as PPARγ and PPARα. These otherbiological activities include, among others, obesity, diabetes, lipidassociated disorders, cancer, inflammatory disorders, disordersinvolving a disrupted or dysfunctional epidermal barrier, and variousother metabolic disorders. Modulators of FXR, both agonists andantagonists, will find utility in treating one or more of thesediseases.

[0005] PCT Publication No. WO 00/40965, which is incorporated herein byreference, describes methods and compositions that are useful formodulating cholesterol levels in a cell and methods for identifyingcompounds that can be tested for ability to modulate cholesterol levelsin mammals. These methods involve analyzing the effect of a testcompound on the binding of FXR to an FXR ligand. Such ligands include,for example, bile acids, coactivators, and corepressors. The methods andcompositions involve modulating FXR-mediated expression of genesinvolved in cholesterol metabolism.

[0006] Despite the advances made by WO 00/40965, there is a need in theart for new FXR modulators, both antagonists and agonists, to be usedfor a variety of indications. The present invention remedies this andother needs.

SUMMARY OF THE INVENTION

[0007] Atherosclerosis is a leading cause of death, myocardialinfarctions, strokes, peripheral vascular disease and cardiovasculardisease. One of the major contributing factors to atherosclerosis ishypercholesteremia. By modulating FXR-mediated expression of genes,using FXR modulating compounds, it is possible to mitigate and therebytreat hypercholesterolemia.

[0008] The present invention provides compounds, pharmaceuticalcompositions and methods that modulate FXR. The invention also providesmethods of using the compounds and compositions for the treatment ofconditions and disorders mediated by FXR, such as atherosclerosis,diabetes, obesity, dyslipidemia, hypercholesterolemia, hypertension,hyperlipidemia and hyperlipoproteinemia, certain inflammatory conditionsand cancer.

[0009] As such, in certain aspects, the present invention providescompounds of Formula I:

B¹-L¹-A¹-L²-B²  I

[0010] In Formula I, A¹ represents a divalent group selected from thefollowing: alkylene, alkenylene, alkynylene, cycloalkylene,cycloalkenylene, arylene, heteroarylene, heterocycloalkylene, andheterocycloalkenylene, or A¹ represents a single or double bond linkingL¹ and L².

[0011] L¹ and L² are each independently selected from the followinggroup of divalent radicals: —O—, —S—, —N(R¹)-, —C(O)—, —C(O)N(R¹)-,—O-alkylene-; —S-alkylene-, —N(R¹)-alkylene, —C(O)-alkylene,—C(O)N(R¹)-alkylene, —C(O)—O-alkylene, alkylene, alkenylene, alkynylene,cycloalkylene, cycloalkenylene, arylene, heteroarylene,heterocycloalkylene, and heterocycloalkenylene.

[0012] B¹ and B² are each independently selected from the group: alkyl,cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycloalkyl, andheterocycloalkenyl.

[0013] In some aspects, L¹ can be additionally linked to B¹ via a groupX¹ to form a 5-9 member ring. In a similar manner, L² can beadditionally linked to B² via a group X² to form a 5-9 member ring.

[0014] X¹ and X² are each independently selected from: a single bond,—O—, —S—, —N(R²)—, —C(O)—, —C(O)N(R²)—, —O-alkylene, —S-alkylene,—N(R²)-alkylene, —C(O)-alkylene, —C(O)N(R²)-alkylene, and—C(O)—O-alkylene.

[0015] R¹ and R² are each independently selected from: hydrogen, alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, aryl(heteroalkyl),(heteroaryl)alkyl, or (heteroaryl)heteroalkyl.

[0016] In another aspect, the present invention provides FXR modulatorsof Formula II:

[0017] In Formula II, A² and A³ are each independently selected from:alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, (heteroaryl)alkyl,aryl(heteroalkyl), or (heteroaryl)heteroalkyl.

[0018] B³ is selected from the following: -hydrogen, -alkylene-C(O)R³,—C(O)R³, alkylene-C(O)N(R³R⁴), —C(O)N(R³R⁴), alkylene-S(O)_(n)N(R³R⁴),—S(O)_(n)N(R³R⁴), alkylene-N(R³R⁴), alkylene-OR³, and —C(O)OR³.

[0019] R³ and R⁴ are each independently selected from: hydrogen, alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, (heteroaryl)alkyl,aryl(heteroalkyl), and (heteroaryl)heteroalkyl.

[0020] X is C, S, or N.

[0021] The subscript p is an integer from 0-2.

[0022] In still another embodiment, the present invention provides FXRmodulators of Formula III:

[0023] In Formula III, A⁴ is selected from the following: —C(O)R⁵,—C(O)N(R⁵R⁶), —S(O)_(n)N(R⁵R⁶), -alkylene-N(R⁵R⁶), -alkylene-OR⁵ and—C(O)OR⁵.

[0024] L³ and L⁴ are each independently selected from the followingdivalent radicals: a single bond, —C(O)—, —S(O)_(p)—, and alkylene,wherein the subscript p is an integer from 0-2.

[0025] B⁴, B⁵ and B⁶ are each independently selected from: alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, fused-benzoheterocycloalkyl,cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, arylalkyl,aryl(heteroalkyl), (heteroaryl)alkyl, and (heteroaryl)heteroalkyl.

[0026] Alternatively, B⁴ and B⁵ join to form a divalent arylene,heteroarylene, alkylene, or cycloalkylene linkage between L³ and L⁴.

[0027] X³ and Y are each independently a trivalent nitrogen atom or atrivalent or tetravalent carbon atom.

[0028] R⁵ and R⁶ are each independently selected from: hydrogen, alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, aryl(heteroalkyl),(heteroaryl)alkyl, and (heteroaryl)heteroalkyl.

[0029] In still yet another aspect, the present invention provides FXRmodulators of Formula IV:

[0030] In Formula IV, A⁵ is selected from the following divalentlinkages: —C(O) -alkylene-, —S(O)_(n)—, —C(O)N(R¹²)-, —S(O)₂N(R¹²)-,-alkylene-N(R¹²)-, -alkylene-O—, or —C(O)O—.

[0031] L⁵ and L⁶ are each independently selected from the followinggroup of divalent radicals: a single bond, —C(O)—, —S(O)_(n)—, andalkylene, wherein the subscript n is an integer from 0-2.

[0032] B⁷, B⁸, and B⁹ are each independently selected from: alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, benzoheterocycloalkyl,cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, arylalkyl,aryl(heteroalkyl), (heteroaryl)alkyl, and (heteroaryl)heteroalkyl.

[0033] Alternatively, B⁷ and B⁸ join to form a divalent arylene,heteroarylene, alkylene, or cycloalkylene linkage between L⁵ and L⁶.

[0034] Z is selected from the following divalent linkages: alkylene,heteroalkylene, cycloalkylene, or heterocycloalkylene.

[0035] X⁷ and Y¹ are independently a trivalent nitrogen atom or atrivalent or tetravalent carbon atom; and

[0036] R¹² is selected from: hydrogen, alkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl,arylalkyl, aryl(heteroalkyl), (heteroaryl)alkyl, and(heteroaryl)heteroalkyl.

[0037] In yet another aspect, the present invention provides FXRmodulators of Formula V:

[0038] In Formula V, A⁶ and A⁷ are each independently selected from:arylene, heteroarylene, cycloalkylene, or heterocycloalkylene.

[0039] B¹⁰ represents: aryl, heteroaryl, arylalkyl, (heteroaryl)alkyl,alkyl, cycloalkyl, cycloalkenyl, heteroalkyl, heterocycloalkyl, orheterocycloalkenyl.

[0040] L⁷ L⁸ and L⁹ are each independently selected from: —O—, —S—,—N(R¹³), —C(O)—, —S(O)—, —S(O)₂—, alkylene, —O-alkylene, —S-alkylene,—N(R¹³)-alkylene, —C(O)-alkylene, —C(O)N(R¹³)-alkylene,—C(O)—O-alkylene, a single bond, or a double bond,

[0041] X⁸ is selected from the following trivalent radicals: N, CR¹³;and

[0042] R¹³ is selected from: hydrogen, alkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl,arylalkyl, or (heteroaryl)alkyl.

[0043] Unless otherwise indicated, the compounds provided in the aboveformula are meant to include pharmaceutically acceptable salts andprodrugs thereof.

[0044] In addition to each of the aspect of the invention providedabove, the present invention further provides pharmaceuticalcompositions containing one or more members of the classes of compoundsabove in admixture with a pharmaceutically acceptable carrier orexcipient. Still further, the invention provides methods of using thecompounds described herein for the treatment of FXR-mediated conditionsand disorders as well as the modulation of cyp7a expression levels inmammals. FXR-mediated conditions and disorders include, but are notlimited to, atherosclerosis, peripheral vascular disease, cardiovasculardisease, hypercholesteremia, cholesterolemia, obesity, diabetes,inflammatory conditions and diseases associated with abnormally high orlow cholesterol levels.

[0045] In other embodiments, the compounds of the present invention areadministered in combination with certain other compounds of the presentinvention “in combination therapy” or in combination with othertherapeutic compounds.

[0046] In yet another embodiment, the present invention provides the useof a compound of Formulae I-V for the manufacture of a medicament fortreatment of an FXR mediated disease or condition.

[0047] These and other aspects will become more apparent when read withthe accompanying diagram and detailed description, which follows.

BRIEF DESCRIPTION OF THE DRAWINGS

[0048] FIGS. 1 (A-E) provide structures of compounds of Formula I.

[0049] FIGS. 2 (A-B) provide structures of compounds of Formula II.

[0050] FIGS. 3 (A-D) provide structures of compounds of Formula III.

[0051] FIGS. 4 (A-C) provide structures of compounds of Formula IV.

[0052]FIG. 5 provides structures of compounds of Formula V.

DETAILED DESCRIPTION OF THE INVENTION AND PREFERRED EMBODIMENTS

[0053] Abbreviations and Definitions

[0054] The abbreviations used herein are conventional, unless otherwisedefined.

[0055] The term “modulate” or “modulation”, as used herein in itsvarious forms, refers to the ability of a compound to activate orinhibit the function of FXR, either directly or indirectly. Modulationmay occur in vitro or in vivo. Modulation, as described herein, isintended to encompass antagonism, agonism, partial antagonism and/orpartial agonism of a function or characteristic associated with FXR.Such compounds are FXR modulating compounds.

[0056] As used herein, the terms “treat”, “treating” and “treatment”includes:

[0057] (1) preventing the disease, i.e. causing the clinical symptoms ofthe disease not to develop in a mammal that may be exposed to orpredisposed to the disease but does not yet experience or displaysymptoms of the disease,

[0058] (2) inhibiting the disease, i.e., arresting or reducing thedevelopment of the disease or its clinical symptoms, or

[0059] (3) relieving the disease, i.e., causing regression of thedisease or its clinical symptoms.

[0060] As used herein, the term “FXR-mediated condition or disorder” andthe like refers to a condition or disorder characterized byinappropriate, e.g., less than or greater than normal, FXR activity.Inappropriate FXR activity might arise as the result of FXR expressionin cells which normally do not express FXR, increased FXR expression ordecreased FXR expression (leading to, e.g., obesity or diabetes).Inappropriate FXR activity might also arise as the result of ligandsecretion by cells which normally do not secrete an FXR ligand,increased FXR ligand expression or decreased FXR ligand expression. AnFXR-mediated condition or disorder may be completely or partiallymediated by inappropriate nuclear receptor activity. However, anFXR-mediated condition or disorder is one in which modulation of FXRresults in some effect on the underlying condition or disease (e.g., anFXR antagonist results in some improvement in patient well-being in atleast some patients).

[0061] As used herein, “obesity” refers to the excessive accumulation ofbody fat. Obesity may have genetic, environmental (e.g., expending lessenergy than is consumed) and regulatory determinants. Cardiovasculardisorders, lipid disorders and metabolic disorders, such ashypertension, hyperlidemia, coronary artery disease and diabetes, arecommonly associated with obesity.

[0062] As used herein, “diabetes” refers to type I diabetes mellitusjuvenile onset diabetes, insulin dependent-diabetes mellitus or IDDM) ortype II diabetes mellitus (non-insulin-dependent diabetes mellitus orNIDDM). The compounds, compositions and methods of the present inventionpreferably are used to treat NIDDM. NIDDM is characterized by insulinresistance and hyperglycemia. Obesity and lipid disorders are commonlyassociated with NIDDM.

[0063] “A therapeutically effective amount” refers to the amount of acompound that, when administered to a mammal for treating a disease, issufficient to effect such treatment for the disease. The“therapeutically effective amount” will vary depending on the compound,the disease and its severity and the age, weight, etc., of the mammal tobe treated.

[0064] The term “alkyl,” by itself or as part of another substituent,means, unless otherwise stated, a straight or branched chain, or cyclichydrocarbon radical, or combination thereof, which may be fullysaturated, mono- or polyunsaturated and can include di- and multivalentradicals, having the number of carbon atoms designated (i.e. C₁-C₁₀means one to ten carbons). Examples of saturated hydrocarbon radicalsinclude groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl,t-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)ethyl,cyclopropylmethyl, homologs and isomers of, for example, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like. An unsaturated alkyl group isone having one or more double bonds or triple bonds. Examples ofunsaturated alkyl groups include vinyl, 2-propenyl, crotyl,2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl),ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs andisomers.

[0065] The term “alkenyl” denotes branched or unbranched hydrocarbonchains containing one or more carbon-carbon double bonds.

[0066] The term “alkynyl” refers to branched or unbranched hydrocarbonchains containing one or more carbon-carbon triple bonds.

[0067] The term “alkylene” by itself or as part of another substituentmeans a divalent radical derived from an alkane, as exemplified by—CH₂CH₂CH₂CH₂—. Typically, an alkylene group will have from 1 to 24carbon atoms, with those groups having 10 or fewer carbon atoms beingpreferred in the present invention. A “lower alkyl” or “lower alkylene”is a shorter chain alkyl or alkylene group, generally having eight orfewer carbon atoms.

[0068] The term “cycloalkylene” by itself or as part of anothersubstituent means a divalent radical derived from an cycloalkane, asexemplified by cyclohexylene. Typically, an cycloalkylene group willhave from 5-8 carbon atoms, with those groups having 6 carbon atomsbeing preferred in the present invention.

[0069] The term “alkenylene” by itself or as part of another substituentmeans a divalent radical derived from an alkenyl, as exemplified by—CH═CHCH₂CH₂—. Typically, alkenylene groups will have from 2 to 24carbon atoms, with those groups having 10 or fewer carbon atoms beingpreferred in the present invention.

[0070] The term “alkynylene” by itself or as part of another substituentmeans a divalent radical derived from an alkynyl, as exemplified by—C≡CCH₂CH₂—. Typically, alkynylene groups will have from 2 to 24 carbonatoms, with those groups having 10 or fewer carbon atoms being preferredin the present invention.

[0071] The terms “alkoxy,” “alkylamino” and “alkylthio” refer to thosegroups having an alkyl group attached to the remainder of the moleculethrough an oxygen, nitrogen or sulfur atom, respectively. Similarly, theterm “dialkylamino” is used in a conventional sense to refer to —NR′R″wherein the R groups can be the same or different alkyl groups.

[0072] The term “heteroalkyl,” by itself or in combination with anotherterm, means, unless otherwise stated, a stable straight or branchedchain, or cyclic hydrocarbon radical, or combinations thereof, fullysaturated or containing from 1 to 3 degrees of unsaturation, consistingof the stated number of carbon atoms and from one to three heteroatomsselected from the group consisting of O, N, Si and S, and wherein thenitrogen and sulfur atoms may optionally be oxidized and the nitrogenheteroatom may optionally be quaternized. The heteroatom(s) O, N and Smay be placed at any interior position of the heteroalkyl group. Theheteroatom Si may be placed at any position of the heteroalkyl group,including the position at which the alkyl group is attached to theremainder of the molecule. Examples include —CH₂—CH₂—O—CH₃,—CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃, —CH₂—CH₂—S(O)—CH₃,—CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃, —Si(CH₃)₃, —CH₂—CH═N—OCH₃, and—CH═CH—N(CH₃)—CH₃. Up to two heteroatoms may be consecutive, such as,for example, —CH₂—NH—OCH₃ and —CH₂—O—Si(CH₃)₃. Also included in the term“heteroalkyl” are those radicals described in more detail below as“heterocycloalkyl.” The term “heteroalkylene” by itself or as part ofanother substituent means a divalent radical derived from heteroalkyl,as exemplified by —CH₂—CH₂—S—CH₂CH₂— and —CH₂—S—CH₂CH₂—NH—CH₂—. Forheteroalkylene groups, heteroatoms can also occupy either or both of thechain termini. Still further, for alkylene and heteroalkylene linkinggroups, no orientation of the linking group is implied.

[0073] The term “acyl” refers to those groups derived from an organicacid by removal of the hydroxy portion of the acid. Accordingly, acyl ismeant to include, for example, acetyl, propionyl, butyryl, decanoyl,pivaloyl, benzoyl and the like.

[0074] An “activated carbonyl” group is a carbonyl group whoseelectrophilicity is enhanced as a result of the groups attached toeither side of the carbonyl. Examples of such activated carbonyl groupsare (polyfluoroalkyl)ketones, (polyfluoroalkyl)aldehydes, alpha-ketoesters, alpha-keto acids, alpha-keto amides, 1,2-diketones,2-acylthiazoles, 2-acylimidazoles, and the like.

[0075] The terms “cycloalkyl” and “heterocycloalkyl”, by themselves orin combination with other terms, represent, unless otherwise stated,cyclic versions of “alkyl” and “heteroalkyl”, respectively.Additionally, for heterocycloalkyl, a heteroatom can occupy the positionat which the heterocycle is attached to the remainder of the molecule.Examples of cycloalkyl include cyclopentyl, cyclohexyl, 1-cyclohexenyl,3-cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkylinclude 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl,tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,1-piperazinyl, 2-piperazinyl, and the like.

[0076] The terms “halo” or “halogen,” by themselves or as part ofanother substituent, mean, unless otherwise stated, a fluorine,chlorine, bromine, or iodine atom. Additionally, terms such as“fluoroalkyl,” are meant to include monofluoroalkyl and polyfluoroalkyl.

[0077] The term “aryl,” employed alone or in combination with otherterms (e.g., aryloxy, arylthioxy, arylalkyl) means, unless otherwisestated, an aromatic substituent which can be a single ring or multiplerings (up to three rings) which are fused together or linked covalently.The term “heteroaryl” is meant to include those aryl rings which containfrom zero to four heteroatoms selected from N, O, and S, wherein thenitrogen and sulfur atoms are optionally oxidized, and the nitrogenatom(s) are optionally quaternized. The “heteroaryl” groups can beattached to the remainder of the molecule through a heteroatom.Non-limiting examples of aryl and heteroaryl groups include phenyl,1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl,3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl,4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl,5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl,2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl,4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl,1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl,3-quinolyl, and 6-quinolyl. Substituents for each of the above notedaryl ring systems are selected from the group of acceptable substituentsdescribed below. The term “arylalkyl” is meant to include those radicalsin which an aryl or heteroaryl group is attached to an alkyl group(e.g., benzyl, phenethyl, pyridylmethyl and the like) or a heteroalkylgroup (e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl,and the like).

[0078] Each of the above terms (e.g., “alkyl,” “heteroalkyl” and “aryl”)are meant to include both substituted and unsubstituted forms of theindicated radical. Preferred substituents for each type of radical areprovided below.

[0079] Substituents for the alkyl and heteroalkyl radicals (includingthose groups often referred to as alkylene, alkenyl, heteroalkylene,heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, andheterocycloalkenyl) can be a variety of groups selected from: —OR′, ═O,═NR′, ═N—OR′, —NR′R″, —SR′, -halogen, -SiR′R″ R′″, —OC(O)R′, —C(O)R′,—CO₂R′, CONR′R″, —OC(O)NR′R″, —NR″C(O)R′, —NR′-C(O)NR″R′″, —NR″C(O)₂R′,—NH—C(NH₂)═NH, —NR′C(NH₂)═NH, —NH—C(NH₂)═NR′, —S(O)R′, S(O)₂R′,—S(O)₂NR′R″, —CN and —NO₂ in a number ranging from zero to (2N+1), whereN is the total number of carbon atoms in such radical. R′, R″ and R′″each independently refer to hydrogen, unsubstituted (C₁-C₈)alkyl andheteroalkyl, unsubstituted aryl, aryl substituted with 1-3 halogens,unsubstituted alkyl, alkoxy or thioalkoxy groups, or aryl-(C₁-C₄)alkylgroups. When R′ and R″ are attached to the same nitrogen atom, they canbe combined with the nitrogen atom to form a 5-, 6-, or 7-membered ring.For example, —NR′R″ is meant to include 1-pyrrolidinyl and4-morpholinyl. From the above discussion of substituents, one of skillin the art will understand that the term “alkyl” is meant to includegroups such as haloalkyl (e.g., —CF₃ and —CH₂CF₃) and acyl (e.g.,—C(O)CH₃, —C(O)CF₃, —C(O)CH₂OCH₃, and the like).

[0080] Similarly, substituents for the aryl groups are varied and areselected from: -halogen, —OR′, —OC(O)R′, —NR′R″, —SR′, —R′, —CN, —NO₂,—CO₂R′, —CONR′R″, —C(O)R′, —OC(O)NR′R″, —NR″C(O)R′, —NR″C(O)₂R′,—NR′-C(O)NR″R′″, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH, —NH—C(NH₂)—NR′, —S(O)R′,—S(O)₂R′, —S(O)₂NR′R″, —NR″-S(O)₂—R′, —N₃, —CH(Ph)₂,perfluoro(C₁-C₄)alkoxy, and perfluoro(C₁-C₄)alkyl, in a number rangingfrom zero to the total number of open valences on the aromatic ringsystem; and where R′, R″ and R′″ are independently selected fromhydrogen, (C₁-C₈)alkyl and heteroalkyl, unsubstituted aryl,(unsubstituted aryl)-(C₁-C₄)alkyl, and (unsubstitutedaryl)oxy-(C₁-C₄)alkyl.

[0081] Two of the substituents on adjacent atoms of the aryl ring mayoptionally be replaced with a substituent of the formula-T-C(O)—(CH₂)_(q)-U-, wherein T and U are independently —NH—, —O—, —CH₂—or a single bond, and the subscript q is an integer of from 0 to 2.Alternatively, two of the substituents on adjacent atoms of the arylring may optionally be replaced with a substituent of the formula-A-(CH₂)_(r)-B-, wherein A and B are independently —CH₂—, —O—, —NH—,—S—, —S(O)—, —S(O)₂—, —S(O)₂NR′- or a single bond, and r is an integerof from 1 to 3. One of the single bonds of the new ring so formed mayoptionally be replaced with a double bond. Alternatively, two of thesubstituents on adjacent atoms of the aryl ring may optionally bereplaced with a substituent of the formula —(CH₂)_(n)—X—(CH₂)_(n)—,where s and t are independently integers of from 0 to 3, and X is —O—,—NR′-, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—. The substituent R′ in —NR′-and —S(O)₂NR′— is selected from hydrogen or unsubstituted (C₁-C₆)alkyl.

[0082] As used herein, the term “heteroatom” is meant to include oxygen(O), nitrogen (N), sulfur (S) and silicon (Si).

[0083] The term “pharmaceutically acceptable salts” is meant to includesalts of the active compounds which are prepared with relativelynontoxic acids or bases, depending on the particular substituents foundon the compounds described herein. When compounds of the presentinvention contain relatively acidic functionalities, base addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable base additionsalts include sodium, potassium, calcium, ammonium, organic amino, ormagnesium salt, or a similar salt. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, oxalic, maleic, malonic, benzoic,succinic, suberic, fumaric, mandelic, phthalic, benzenesulfonic,p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Alsoincluded are salts of amino acids such as arginate and the like, andsalts of organic acids like glucuronic or galactunoric acids and thelike (see, for example, Berge et al. (1977) J. Pharm. Sci. 66:1-19).Certain specific compounds of the present invention contain both basicand acidic functionalities that allow the compounds to be converted intoeither base or acid addition salts.

[0084] The neutral forms of the compounds may be regenerated bycontacting the salt with a base or acid and isolating the parentcompound in the conventional manner. The parent form of the compounddiffers from the various salt forms in certain physical properties, suchas solubility in polar solvents, but otherwise the salts are equivalentto the parent form of the compound for the purposes of the presentinvention.

[0085] In addition to salt forms, the present invention providescompounds, which are in a prodrug form. Prodrugs of the compoundsdescribed herein are those compounds that readily undergo chemicalchanges under physiological conditions to provide the compounds of thepresent invention. Additionally, prodrugs can be converted to thecompounds of the present invention by chemical or biochemical methods inan ex vivo environment. For example, prodrugs can be slowly converted tothe compounds of the present invention when placed in a transdermalpatch reservoir with a suitable enzyme or chemical reagent. Prodrugs areoften useful because, in some situations, they may be easier toadminister than the parent drug. They may, for instance, be bioavailableby oral administration whereas the parent drug is not. The prodrug mayalso have improved solubility in pharmacological compositions over theparent drug. Certain compounds of the present invention can exist inunsolvated forms as well as solvated forms, including hydrated forms. Ingeneral, the solvated forms are equivalent to unsolvated forms and areintended to be encompassed within the scope of the present invention.Certain compounds of the present invention may exist in multiplecrystalline or amorphous forms. In general, all physical forms areequivalent for the uses contemplated by the present invention and areintended to be within the scope of the present invention.

[0086] Certain compounds of the present invention possess asymmetriccarbon atoms (optical centers) or double bonds; the racemates,diastereomers, geometric isomers and individual isomers are all intendedto be encompassed within the scope of the present invention.

[0087] The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areintended to be encompassed within the scope of the present invention.

[0088] Compounds

[0089] In one aspect, the present invention provides FXR modulators ofFormula I:

B¹-L¹-A¹-L²-B²  I

[0090] In Formula I, A¹ represents a divalent group selected from thefollowing: alkylene, alkenylene, alkynylene, cycloalkylene,cycloalkenylene, arylene, heteroarylene, heterocycloalkylene, andheterocycloalkenylene, or A¹ represents a single or double bond linkingL¹ and L². Preferably, A¹ is (C₁-C₈)alkylene, arylene (e.g., phenylene)or heteroarylene or a single bond. More preferably, A¹ is(C₁-C₆)alkylene, phenylene or a divalent pyridine group. Even morepreferably, A¹ is (C₁-C₄)alkylene, arylene, or a divalent pyridinegroup.

[0091] L¹ and L² are each independently selected from the followinggroup of divalent radicals: —O—, —S—, —N(R¹)-, —C(O)—, —C(O)N(R¹)-,—O-alkylene-, —S-alkylene-, —N(R¹)-alkylene, —C(O)-alkylene,—C(O)N(R¹)-alkylene, —C(O)—O-alkylene, alkylene, alkenylene, alkynylene,cycloalkylene, cycloalkenylene, arylene, heteroarylene,heterocycloalkylene, and heterocycloalkenylene. In the above alkyleneand alkylene-containing groups, the number of carbon atoms willtypically be from one to eight, and preferably, one to four carbonatoms. Cycloalkylene and unsaturated forms thereof, will typicallycontain four to seven carbon atoms, with those containing five or sixcarbon atoms being preferred.

[0092] B¹ and B² are each independently selected from the group: alkyl,cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycloalkyl, andheterocycloalkenyl.

[0093] In some aspects, L¹ can be additionally linked to B¹ via a groupX¹ to form a 5-9 member ring. In a similar manner, L² can beadditionally linked to B² via a group X² to form a 5-9 member ring. Inpreferred embodiments, B¹ and B² are independently selected from:(C₁-C₈)alkyl, (C₅-C₈)cycloalkyl, (C₅-C₈)cycloalkenyl, aryl, heteroaryl,aryl(C₁-C₄)alkyl, (heteroaryl)(C₁-C₄)alkyl, and a five to eight-memberedheterocycloalkyl group. Preferably, B¹ and B² are each independently asubstituted or unsubstituted phenyl group, a substituted orunsubstituted benzyl group or a (C₅-C₈)cycloalkyl group. In the mostpreferred embodiments, B¹ and B² are the same.

[0094] R¹ and R² are each independently selected from: hydrogen, alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, aryl(heteroalkyl),(heteroaryl)alkyl, or (heteroaryl)heteroalkyl. More preferably, R¹ andR² are each (C₁-C₈)alkyl, (C₁-C₈)heteroalkyl, (C₁-C₈)cycloalkyl,(C₁-C₈)heterocycloalkyl, aryl, heteroaryl, aryl(C₁-C₄)alkyl, or(heteroaryl)(C₁-C₄)alkyl. Still more preferably, R¹ and R² aresubstituted or unsubstituted phenyl group, a substituted orunsubstituted benzyl group, a (C₅-C₈)cycloalkyl group or a (C₁-C₈)alkylgroup.

[0095] X¹ and X² are each independently selected from: a single bond,—O—, —S—, —N(R²)—, —C(O)—, —C(O)N(R²)—, —O-alkylene, —S-alkylene,—N(R²)-alkylene, —C(O)-alkylene, —C(O)N(R²)-alkylene, and—C(O)—O-alkylene. Preferably, X¹ and X² are each independently a(C₁-C₈)alkylene group, with (C₁-C₃)alkylene groups being more preferred.

[0096] In certain aspects, the preferred compounds of Formula I arethose in which:

[0097] A¹ is (C₁-C₈)alkylene, arylene, heteroarylene or a single bond.

[0098] L¹ and L² are —C(O)— or —C(O)N(R¹)-.

[0099] R¹ is (C₅-C₈)cycloalkyl, aryl, heteroaryl, aryl(C₁-C₄)alkyl, or(heteroaryl)(C¹-C₄)alkyl; and

[0100] B¹ and B² are aryl, heteroaryl, aryl(C₁-C₄)alkyl,(heteroaryl)(C₁-C₄)alkyl, (C₁-C₈)alkyl, or (C₅-C⁸)cycloalkyl.

[0101] In other aspects, preferred compounds of Formula I are those inwhich A¹ is (C₁-C₈)alkylene, phenylene, divalent pyridine or a singlebond.

[0102] L¹ and L² are —C(O)— or —C(O)N(R¹)-.

[0103] R¹ is substituted or unsubstituted (C₅-C₈)cycloalkyl, substitutedor unsubstituted phenyl, substituted or unsubstituted benzyl, or(C₁-C₈)alkyl; and

[0104] B¹ and B² are independently substituted or unsubstituted(C₅-C₈)cycloalkyl, substituted or unsubstituted phenyl, substituted orunsubstituted benzyl. More preferably, B¹ and B² are the same.

[0105] In yet other aspects, preferred compounds of Formula I are thosein which:

[0106] A¹ is alkylene, arylene, heteroarylene or a single bond.

[0107] L¹ and L² are —C(O)N(R¹)-.

[0108] R¹ is aryl, heteroaryl, arylalkyl, or (heteroaryl)alkyl.

[0109] B¹ and B² are each independently aryl, heteroaryl, arylalkyl,(heteroaryl)alkyl, alkyl, or cycloalkyl.

[0110] In another preferred aspect, compounds of Formula I are those inwhich:

[0111] A¹ is heteroarylene containing two fused rings.

[0112] L¹ and L² are —O—, —NH—, or —N(R¹)-.

[0113] R¹ is alkyl or heteroalkyl; and

[0114] B¹ and B² are aryl, heteroaryl, arylalkyl, (heteroaryl)alkyl,alkyl, or cycloalkyl. Compounds of Formula I act primarily as FXRantagonists, but in certain instances, the compounds act as FXRagonists. Preferred compounds of Formula I are set forth in FIGS. 1A-1E.

[0115] In another aspect, the present invention provides FXR modulatorsof Formula II:

[0116] In Formula II, A² and A³ are each independently selected from:alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, (heteroaryl)alkyl,aryl(heteroalkyl), or (heteroaryl)heteroalkyl.

[0117] B³ is selected from the following: hydrogen, -alkylene-C(O)R³,—C(O)R³, alkyklene-C(O)N(R³R⁴), —C(O)N(R³R⁴), alkylene-S(O)_(n)N(R³R⁴),—S(O)_(n)N(R³R⁴), alkylene-N(R³R⁴), alkylene-OR³, and —C(O)OR³.

[0118] R³ and R⁴ are each independently selected from: hydrogen, alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, (heteroaryl)alkyl,aryl(heteroalkyl), and (heteroaryl)heteroalkyl.

[0119] X is C, S, or N.

[0120] The subscripts n and p are each independently an integer from0-2, provided that the following compound is excluded:

[0121] In one aspect, preferred compounds of Formula II are those inwhich: A² is an aryl group substituted ortho to the nitrogen withhydrogen bond donating groups including, but not limited to: —OH, —NH₂,—NHC(O)-alkyl, —NHSO₂-alkyl.

[0122] A³ is aryl or heteroaryl.

[0123] B³ is hydrogen.

[0124] X is C; and

[0125] p is 1.

[0126] In another aspect, preferred compounds of Formula II are those inwhich:

[0127] A² and A³ are aryl or heteroaryl.

[0128] B³ is alkylene-C(O)N(R³R⁴), or alkylene-S(O)_(n)N(R³R⁴),

[0129] wherein R³ is arylalkyl or (heteroaryl)alkyl and R⁴ is hydrogen

[0130] X is S; and

[0131] n is 2.

[0132] Compounds of Formula II act primarily as FXR antagonists, but incertain instances, act as FXR agonists. Preferred compounds of FormulaII are set forth in FIGS. 2A-2-B.

[0133] In still another aspect, the present invention provides FXRmodulators of Formula III:

[0134] In Formula III, A⁴ is selected from the following: hydrogen,—C(O)R⁵, —C(O)N(R⁵R⁶), —S(O)_(n)N(R⁵R⁶), -alkylene-N(R⁵R⁶),-alkylene-OR⁵, and —C(O)OR⁵.

[0135] L³ and L⁴ are each independently selected from the followingdivalent radicals: a single bond, —C(O)—, —S(O)_(p)—, and alkylene,wherein the subscript p is an integer from 0-2.

[0136] B⁴, B⁵ and B⁶ are each independently selected from: hydrogen,alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,fused-benzoheterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl,heteroaryl, arylalkyl, aryl(heteroalkyl), (heteroaryl)alkyl, and(heteroaryl)heteroalkyl.

[0137] Alternatively, B⁴ and B⁵ join to form a divalent arylene,heteroarylene, alkylene, or cycloalkylene linkage between L³ and L⁴, B⁶is hydrogen, alkyl, heteroalkyl, heterocycloalkyl, arylalkyl or(heteroaryl)alkyl.

[0138] X³ and Y are independently a trivalent nitrogen atom or atrivalent or tetravalent carbon atom.

[0139] R⁵ and R⁶ are each independently selected from: hydrogen, alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, aryl(heteroalkyl),(heteroaryl)alkyl, and (heteroaryl)heteroalkyl.

[0140] In certain aspects, preferred compounds of Formula III are thosein which

[0141] A⁴ is hydrogen, —C(O)N(R⁵R⁶) or —S(O)₂N(R⁵R⁶).

[0142] R⁵ and R⁶ are each independently alkyl, cycloalkyl, orheterocycloalkyl.

[0143] L³ and L⁴ are each independently —C(O)—, —S(O)₂—, or loweralkylene.

[0144] B⁴ and B⁵ join to form an arylene or heteroarylene linkagebetween L³ and L⁴.

[0145] X is tetravalent carbon in the R configuration.

[0146] Y is trivalent nitrogen; and

[0147] B⁶ is hydrogen, alkyl, heteroalkyl, heterocycloalkyl, arylalkyl,or (heteroaryl)alkyl.

[0148] In another preferred aspect, compounds of Formula III are thosein which:

[0149] A⁴ is hydrogen, —C(O)N(R⁵R⁶) or —S(O)₂N(R⁵R⁶).

[0150] R⁵ and R⁶ are each independently alkyl, cycloalkyl, orheterocycloalkyl.

[0151] L³ and L⁴ are independently —C(O)—, —S(O)₂—, or lower alkylene.

[0152] B⁴ and B⁵ are independently hydrogen, alkyl, arylalkyl, aryl, orheteroaryl.

[0153] X is tetravalent carbon in the R configuration.

[0154] Y is trivalent nitrogen; and

[0155] B⁶ is hydrogen, alkyl, heteroalkyl, heterocycloalkyl, arylalkyl,or (heteroaryl)alkyl. Compounds of Formula III act primarily as FXRantagonists, but in certain instances, act as FXR agonists. Preferredcompounds of Formula III are set forth in FIGS. 3A-3D.

[0156] In another embodiment, the present invention provides compoundsof Formula IIIa.

[0157] wherein:

[0158] X⁴, X⁵ and X⁶ are each independently C or S.

[0159] R¹⁰ and R¹ are each independently alkyl, cycloalkyl, orheterocycloalkyl.

[0160] R⁹ is an optionally substituted aryl, heteroaryl, arylalkyl,(heteroaryl)alkyl, heterocycloalkyl; and the subscripts p, q, and r areeach independently integers from 0-2.

[0161] R¹⁴ is selected from hydrogen, halogen, alkyl, alkoxy,alkylamino, alkylthio, acyl, cycloalkyl and aryl.

[0162] In yet another embodiment, the present invention providescompounds of Formula IIIb.

[0163] In Formula IIIb, A⁴ is selected from hydrogen, —C(O)R⁵,—C(O)N(R⁵R⁶), —S(O)_(n)N(R⁵R⁶), -alkylene-N(R⁵R⁶), -alkylene-OR⁵ and—C(O)OR⁵.

[0164] B⁵ and B⁶ are independently selected from hydrogen, alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, fused-benzoheterocycloalkyl,cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, arylalkyl,aryl(heteroalkyl), (heteroaryl)alkyl and (heteroaryl)heteroalkyl.

[0165] X³ is a trivalent nitrogen atom or a trivalent or tetravalentcarbon atom.

[0166] In still yet another aspect, the present invention provides FXRmodulators of Formula IV:

[0167] In Formula IV, A⁵ is selected from the following divalentlinkages: —C(O)—, -alkylene-, —S(O)_(n)—, —C(O)N(R¹²)—, —S(O)₂N(R¹²)—,-alkylene-N(R²)-, -alkylene-O—, or —C(O)O—.

[0168] L⁵ and L⁶ are each independently selected from the followinggroup of divalent radicals: a single bond, —C(O)—, —S(O)_(n)—, andalkylene, wherein the subscript n is an integer from 0-2.

[0169] B⁷, B⁸, and B⁹ are each independently selected from: alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, benzoheterocycloalkyl,cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, arylalkyl,aryl(heteroalkyl), (heteroaryl)alkyl, and (heteroaryl)heteroalkyl.

[0170] Alternatively, B⁷ and B⁸ join to form a divalent arylene,heteroarylene, alkylene, or cycloalkylene linkage between L⁵ and L⁶.

[0171] Z is selected from the following divalent linkages: alkylene,heteroalkylene, cycloalkylene, or heterocycloalkylene.

[0172] X⁷ and Y¹ are independently a trivalent nitrogen atom or atrivalent or tetravalent carbon atom; and

[0173] R¹² is selected from: hydrogen, alkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl,arylalkyl, aryl(heteroalkyl), (heteroaryl)alkyl, and(heteroaryl)heteroalkyl.

[0174] In certain aspects, preferred compounds of Formula IV are thosein which:

[0175] A⁵ is —C(O)—, —C(O)N(R¹²)- or —S(O)₂N(R¹²)-.

[0176] R¹² is alkyl, cycloalkyl, or heterocycloalkyl.

[0177] B⁷ and B⁸ are combined in an arylene or heteroarylene linkagebetween L⁵ and L⁶.

[0178] B⁹ is alkyl, heteroalkyl, heterocycloalkyl, arylalkyl, or(heteroaryl)alkyl.

[0179] Z is alkylene, heteroalkylene, or heterocycloalkylene.

[0180] L⁵ and L⁶ are independently —C(O)—, —S(O)₂—, or lower alkylene.

[0181] X⁷ is tetravalent carbon; and

[0182] Y¹ is trivalent nitrogen. Compounds of Formula IV act primarilyas FXR antagonists, but some of the compounds act as FXR agonists.Preferred compounds of Formula IV are set forth in FIGS. 4A-4C.

[0183] In yet another aspect, the present invention provides novel FXRmodulators of Formula V:

[0184] In Formula V, A⁶ and A⁷ are each independently selected from:arylene, heteroarylene, cycloalkylene, or heterocycloalkylene.

[0185] B¹⁰ represents: aryl, heteroaryl, arylalkyl, (heteroaryl)alkyl,alkyl, cycloalkyl, cycloalkenyl, heteroalkyl, heterocycloalkyl, orheterocycloalkenyl.

[0186] L⁷ L⁸ and L⁹ are each independently selected from: —O—, —S—,—N(R¹³), —C(O)—, —S(O)—, —S(O)₂—, alkylene, —O-alkylene, —S-alkylene,—N(R¹³)-alkylene, —C(O)—alkylene, —C(O)N(R¹)-alkylene, —C(O)—O-alkylene,a single bond, or a double bond;

[0187] X⁸ is selected from the following trivalent radicals: N, CR¹³;and

[0188] R¹³ is selected from: hydrogen, alkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl,arylalkyl, or (heteroaryl)alkyl.

[0189] In certain aspects, A⁶ and A⁷ are aryl, heteroaryl, cycloalkyl,or heterocycloalkyl;

[0190] B¹⁰ is aryl, heteroaryl, arylalkyl, or (heteroaryl)alkyl;

[0191] L⁷ and L⁸ are independently selected from —C(O)—, —S(O)—, or—S(O)₂—;

[0192] L⁹ is —C(O)—, alkylene, or a single bond; and

[0193] X⁵ is N. Compounds of Formula V act primarily as FXR agonists,but in certain aspects act as FXR antagonists. Preferred compounds ofFormula V are set forth in FIG. 5.

[0194] The vast majority of the compounds contemplated for use in thepresent invention are novel, while one or more of the specific compoundsset forth in FIGS. 1-5 may be commercially available. The presentinvention specifically contemplates the exclusion, by way of proviso, ofcommercially available compounds from the compound claims (and, ifappropriate, from the pharmaceutical composition claims). Unlessotherwise indicated, it is to be understood that the present inventionincludes those compounds that are novel. The pharmaceuticalcompositions, various methods (e.g., methods of treating certainFXR-mediated conditions and disorders), and the like include both thenovel compounds of the invention and compounds that are commerciallyavailable.

[0195] Synthetic Schemes

[0196] Illustrative synthetic methods for representative compounds inFormulae I-V are provided in the examples set forth below. One of skillin the art will appreciate that substitution of one starting material orreagent for another, will provide additional compounds within the scopeof the present invention. For example, in Example I the synthesis ofcompound 1.1.4 is set forth. A skilled artisan will readily appreciatehowever, that isophthaloyl dichloride can be reacted withN-benzyl-p-anisidine as indicated, or it can be reacted with a varietyof other substituted N-benzyl anilines.

[0197] In a similar manner, substitution of isophthaloyl dichloride with2,6-pyridinedicarbonylchloride (commercially available from AldrichChemical Co., Milwaukee, Wis. USA) will provide compounds of Formula Iwherein A¹ is a heteroarylene group. A number of symmetrical compoundsof Formula I are provided in FIGS. 1A-1E. All of these compounds can beprepared by analogous amide bond-forming reactions using commerciallyavailable starting materials or materials that are readily availablefrom established literature procedures.

[0198] Further examples of compounds of Formula I and their synthesesare set forth in Examples 2 and 3. In Example 3, a2,3-dichloro-6-nitroquinoxaline is reacted with an excess (over twoequivalents) of a substituted aniline to provide the target compound.Similarly, other substituted 2,3-dichloroquinoxalines can be used withother substituted anilines, arylalkylamines or alkylamines.Unsymmetrically substituted compounds can be prepared as described forcompound 1.2.26 of FIG. 1D, replacing 3,4-dimethylaniline with othersubstituted anilines and replacing cyclohexylamine with otheralkylamines, cycloalkylamines, arylalkylamines and anilines.

[0199] A representative compound of Formula II is set forth in Example4. As illustrated therein, N-(2,6-dihydoxy-phenyl)-benzamide wassuspended in 85% H₃PO₄ and 2-methyl-2-propanol. The reaction mixture wasvigorously stirred and then poured into of deionized water and extractedwith ethyl acetate to generate compound 2.1.1 (see, FIG. 2A).

[0200] A further example of a compound of Formula II is shown in Example5. As illustrated therein,N-(3-trifluoromethyl-phenyl)-benzenesulfonamide is dissolved inanhydrous THF and then treated with potassium tert-butoxide and achloroacetamide. Thereafter, Bu₄NI was added to catalyze the alkylationreaction. After work up, compound 2.2.10 was produced in good yield.Those of skill in the art will appreciate that suitable modifications ofExample 4 and 5 will generate further compounds of Formula II.

[0201] Likewise, compounds of Formula III can be prepared as set forthin Example 6, wherein the synthesis of compound 3.1.1 is illustrated. Askilled artisan will appreciate that by suitable modification of Example6, as for example, replacing N-phthaloyl-DL-phenylalanine by a varietyof N-phthaloyl-DL-amino acids, additional compounds of the invention canbe generated. Dicyclohexylamine can also be replaced with otherdicycloalkylamines, N-cylcoalkyl anilines, N-cycloalkyl N-benzylamines,and the like.

[0202] Compounds of Formula IV can be prepared using methods similar tothose described in Example 8, wherein the synthesis of compound 4.7 isillustrated. In this example, N-substituted amino acids (N-benzyl,N-benzoyl, N-alkyl, N,N-dialkyl and the like) can be converted to amidesof Formula IV by simple coupling reactions using, for example,dicyclohexylamine, isopropyl t-butyl amine, 3-pentyl t-butyl amine, andthe like. Other examples of compounds of Formula IV are set forth inFIG. 4A-4C.

[0203] In addition, compounds of Formula V can be prepared using methodssimilar to those described in Example 9, wherein the synthesis of 5.3 isillustrated. As shown therein, an ortho-nitro diaryl sulfonamide (e.g.,compound 5.3a in Example 9) can be converted to an ortho amino compound(see, for example, compound 5.3b in Example 9) then oxidized to adiazonium compound that undergoes an intramolecular C—H bond insertionon the nearby phenyl ring to form a tricyclic structure (e.g., 5.3c inExample 9). Sulfonylation of the sulfonamide nitrogen of 5.3c can beaccomplished using standard conditions. Alternatively, the nitrogen canbe acylated with, for example, benzoyl chloride or a substituted benzoylchloride to form other compounds useful as FXR antagonists. Forcompounds that are symmetrical, N-phthaloyl amino acids can be reactedwith, for example, diamines such as 1,6-hexanediamine, N,N′-dimethylbutanediamine, 4,13-diaza-18-crown-6, 7,13-diaza-15-crown-5,N,N′-dimethyl 2-aminoethyl ether, 1,4-bis(aminomethyl)cyclohexane, andthe like.

[0204] Compositions

[0205] In view of the FXR antagonism and in some cases, agonism effectsof the compounds described herein, the present invention furtherprovides pharmaceutical compositions comprising one or more of the abovecompounds in combination with a pharmaceutically acceptable carrier orexcipient.

[0206] In one embodiment, the present invention provides the subjectcompounds combined with a pharmaceutically acceptable excipient such assterile saline or other medium, water, gelatin, an oil, etc. to formpharmaceutically acceptable compositions. The compositions and/orcompounds may be administered alone or in combination with anyconvenient carrier, diluent, etc. and such administration may beprovided in single or multiple dosages. Useful carriers include, but arenot limited to, solid, semi-solid or liquid media including water andnon-toxic organic solvents.

[0207] In another embodiment, the present invention provides the subjectcompounds in the form of a prodrug, which can be metabolically orchemically converted to the subject compounds by the recipient host. Awide variety of prodrug derivatives are known in the art, such as thosethat rely on hydrolytic cleavage or oxidative activation of the prodrug.An example, without limitation, of a prodrug would be a compound of thepresent invention which is administered as an ester (the “prodrug”), butthen is metabolically hydrolyzed to the carboxylic acid, the activeentity. Additional examples include peptidyl derivatives of a compoundof the invention.

[0208] The compositions can be provided in any convenient form,including tablets, capsules, lozenges, troches, hard candies, powders,sprays, creams, suppositories, etc. As such, the compositions, inpharmaceutically acceptable dosage units or in bulk, may be incorporatedinto a wide variety of containers. For example, dosage units may beincluded in a variety of containers including capsules, pills, etc.

[0209] The pharmaceutical compositions of the present invention aresuitable for use in a variety of drug delivery systems. Examples ofsuitable formulations for use in the present invention are found inRemington's Pharmaceutical Sciences (Mack Publishing Company,Philadelphia, Pa., 17th ed. (1985)), which is incorporated herein byreference. In addition, for a brief review of methods for drug delivery,see, Langer, Science 249:1527-1533 (1990), which is incorporated hereinby reference.

[0210] The pharmaceutical compositions of the present invention areintended for parenteral, topical, oral or local administration. Incertain aspects, the pharmaceutical compositions are administeredparenterally, e.g., intravenously, subcutaneously, intradermally, orintramuscularly. In one embodiment, the invention provides compositionsfor parenteral administration which comprise a compound of the presentinvention, dissolved or suspended in an acceptable carrier, preferablyan aqueous carrier. A variety of aqueous carriers may be used including,for example, water, buffered water, 0.4% saline, 0.3% glycine,hyaluronic acid and the like. These compositions may be sterilized byconventional, well-known sterilization techniques or, they may besterile filtered. The resulting aqueous solutions may be packaged foruse as is or lyophilized, the lyophilized preparation being combinedwith a sterile solution prior to administration. The compositions maycontain pharmaceutically acceptable auxiliary substances as required toapproximate physiological conditions including pH adjusting andbuffering agents, tonicity adjusting agents, wetting agents and thelike, such as, for example, sodium acetate, sodium lactate, sodiumchloride, potassium chloride, calcium chloride, sorbitan monolaurate,triethanolamine oleate, etc.

[0211] For solid formulations, compounds of the present invention can beadmixed with conventional nontoxic solid carriers can be used whichinclude, for example, pharmaceutical grades of mannitol, lactose,starch, magnesium stearate, sodium saccharin, talcum, cellulose,glucose, sucrose, magnesium carbonate, and the like. For oraladministration, a pharmaceutically acceptable nontoxic composition isformed by incorporating any of the normally employed carriers orexcipients, such as those carriers previously listed, and generally10-95% of active ingredient and more preferably at a concentration of25%-75%.

[0212] For aerosol administration, the compounds of the presentinvention and antidiabetic agents are preferably supplied in finelydivided form along with a surfactant and propellant. The surfactantmust, of course, be nontoxic, and preferably soluble in the propellant.Representative of such agents are the esters or partial esters of fattyacids containing from 6 to 22 carbon atoms, such as caproic, octanoic,lauric, palmitic, stearic, linoleic, linolenic, olesteric and oleicacids with an aliphatic polyhydric alcohol or its cyclic anhydride.Mixed esters, such as mixed or natural glycerides may be employed. Acarrier can also be included, as desired, as with, e.g., lecithin forintranasal delivery.

[0213] The compounds of the present invention can be prepared andadministered in a wide variety of oral and parenteral dosage forms.Thus, the compounds of the present invention can be administered byinjection, that is, intravenously, intramuscularly, intracutaneously,subcutaneously, intraduodenally, or intraperitoneally. Also, thecompounds described herein can be administered by inhalation, forexample, intranasally. Additionally, the compounds of the presentinvention can be administered transdermally. Accordingly, the presentinvention also provides pharmaceutical compositions comprising apharmaceutically acceptable carrier or excipient and either a compoundof the present invention or a pharmaceutically acceptable salt. Thepresent invention also contemplates the use of depot formulations.

[0214] For preparing pharmaceutical compositions from the compounds ofthe present invention, pharmaceutically acceptable carriers can beeither solid or liquid. Solid form preparations include powders,tablets, pills, capsules, cachets, suppositories, and dispersiblegranules. A solid carrier can be one or more substances that may alsoact as diluents, flavoring agents, binders, preservatives, tabletdisintegrating agents, or an encapsulating material.

[0215] In powders, the carrier is a finely divided solid that is in amixture with the finely divided active component. In tablets, the activecomponent is mixed with the carrier having the necessary bindingproperties in suitable proportions and compacted in the shape and sizedesired.

[0216] The powders and tablets preferably contain from 5% to 70% of theactive compound. Suitable carriers are magnesium carbonate, magnesiumstearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin,tragacanth, methylcellulose, sodium carboxymethylcellulose, a lowmelting wax, cocoa butter, and the like. The term “preparation” isintended to include the formulation of the active compound withencapsulating material as a carrier providing a capsule in which theactive component with or without other carriers, is surrounded by acarrier, which is thus in association with it. Similarly, cachets andlozenges are included. Tablets, powders, capsules, pills, cachets, andlozenges can be used as solid dosage forms suitable for oraladministration.

[0217] For preparing suppositories, a low melting wax, such as a mixtureof fatty acid glycerides or cocoa butter, is first melted and the activecomponent is dispersed homogeneously therein, as by stirring. The moltenhomogeneous mixture is then poured into convenient sized molds, allowedto cool, and thereby to solidify.

[0218] Liquid form preparations include solutions, suspensions, andemulsions, for example, water or water/propylene glycol solutions. Forparenteral injection, liquid preparations can be formulated in solutionin aqueous polyethylene glycol solution.

[0219] Aqueous solutions suitable for oral use can be prepared bydissolving the active component in water and adding suitable colorants,flavors, stabilizers, and thickening agents as desired. Aqueoussuspensions suitable for oral use can be made by dispersing the finelydivided active component in water with viscous material, such as naturalor synthetic gums, resins, methylcellulose, sodiumcarboxymethylcellulose, and other well-known suspending agents.

[0220] Also included are solid form preparations that are intended to beconverted, shortly before use, to liquid form preparations for oraladministration. Such liquid forms include solutions, suspensions, andemulsions. These preparations may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents, andthe like.

[0221] The pharmaceutical preparation is preferably in unit dosage form.In such form the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

[0222] The quantity of active component in a unit dose preparation maybe varied or adjusted from 0.1 mg to 1000 mg, preferably 1.0 mg to 100mg according to the particular application and the potency of the activecomponent. The composition can, if desired, also contain othercompatible therapeutic agents.

[0223] It will be appreciated that the actual preferred course oftherapy will vary according to, inter alia, the mode of administrationof the compound of the present invention, the particular formulationbeing utilized, the mode of administration of the compounds, theparticular disease being treated and the particular host being treated.The optimal course of therapy for a given set of conditions can beascertained by those skilled in the art using conventional course oftherapy determination tests and in view of the information set outherein.

[0224] Methods and Uses, Dosages and Schedules

[0225] In another embodiment, the present invention provides methods ofusing the compounds and pharmaceutical compositions described herein forthe treatment of FXR modulated diseases and conditions. In certainaspects, the compounds of the present invention that are administeredcomprise a compound of Formulae I-V formulated individually, together,or with one or more additional active agents. In other embodiments, thepresent invention provides the use of a compound of Formulae I-V for themanufacture of a medicament for treatment of an FXR mediated disease orcondition.

[0226] In therapeutic use for the treatment of obesity, atherosclerosis,peripheral vascular disease, hypercholesteremia, diabetes, orinflammatory conditions, the compounds utilized in the pharmaceuticalmethod of the invention are administered at the initial dosage of about0.001 mg/kg to about 100 mg/kg daily. A daily dose range of about 0.1mg/kg to about 10 mg/kg is preferred. The dosages, however, may bevaried depending upon the requirements of the patient, the severity ofthe condition being treated, and the compound being employed.Determination of the proper dosage for a particular situation is withinthe skill of the practitioner. Generally, treatment is initiated withsmaller dosages that are less than the optimum dose of the compound.Thereafter, the dosage is increased by small increments until theoptimum effect under circumstances is reached. For convenience, thetotal daily dosage may be divided and administered in portions duringthe day, if desired.

[0227] In therapeutic applications, the compounds of the presentinvention are administered to a patient in a amount sufficient to elicita response. An amount adequate to accomplish this is defined as“therapeutically effective combination dose.”

[0228] Effective combination amounts for various uses will depend on,for example, the particular compound of the present invention employed,the manner of administration, the weight and general state of health ofthe patient, and the judgment of the prescribing physician. In oneembodiment, the composition or formulation to be administered willcontain a quantity of a compound(s) according to Formulae I-V in anamount effective to treat the disease/condition of the subject beingtreated.

[0229] In certain instances, administration of the compounds of thepresent invention can be via any method, which provides systemicexposure to the compound of this invention, preferably to the muscle andfatty tissue. These methods include oral routes, parenteral,intraduodenal routes, etc. Generally, the compounds of the presentinvention are administered in single (e.g., once daily) or multipledoses. The compounds of the present invention are generally administeredin the form of a pharmaceutical composition comprising at least one ofthe compounds of Formulae I-V together with a pharmaceuticallyacceptable carrier or diluent. Thus, the compounds of this invention canbe administered individually or together in any conventional oral,parenteral or transdermal dosage form.

[0230] For oral administration, a pharmaceutical composition can takethe form of solutions, suspensions, tablets, pills, capsules, powders,and the like. Tablets containing various excipients such as sodiumcitrate, calcium carbonate and calcium phosphate are employed along withvarious binders such as starch and preferably potato or tapioca starchand certain complex silicates, together with binding agents such aspolyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally,lubricating agents such as magnesium stearate, sodium lauryl sulfate andtalc are often very useful for tabletting purposes. Solid compositionsof a similar type are also employed as fillers in soft and hard-filledgelatin capsules; preferred materials in this connection also includelactose or milk sugar as well as high molecular weight polyethyleneglycols. When aqueous suspensions and/or elixirs are desired for oraladministration, the compounds of this invention can be combined withvarious sweetening agents, flavoring agents, coloring agents,emulsifying agents and/or suspending agents, as well as such diluents aswater, ethanol, propylene glycol, glycerin and various like combinationsthereof.

[0231] In other aspects, the compounds and methods of the presentinvention are useful in treating various skin conditions and skinailments. Skin conditions that are treatable by the compounds of thisinvention include, but are not limited to, the skin of premature infantsof gestational age less than 33 weeks; atopic and seborrheic dermatitis;inflammation to mucous membranes, such as cheilitis, chapped lips, nasalirritation and vulvovaginitis; eczematous dermatitis resulting fromallergic and irritant contact, eczema craquelee, radiation and stasisdermatitis; ulcers and erosions due to chemical or thermal burns,bullous disorders, or vascular compromise or ischemia including venous,arterial, embolic or diabetic ulcers; ichthyoses, with or without anassociated barrier abnormality; epidermolysis bullosa; psoriasis;hypertrophic scars and keloids; intrinsic aging and/or dermatoheliosus;mechanical friction blistering; corticosteroid atrophy; and melanoma andnon-melanoma skin cancer, including lignin melanoma, basal cellcarcinoma, squamous cell carcinoma, actinic keratoses, and virallyinduced neoplasia (warts and condylomata accuminata) (see, U.S. Pat. No.6,060,515, incorporated herein by reference).

[0232] In certain embodiments, the compounds of the present inventionare administered in combination with certain other compounds of thepresent invention “in combination therapy” or in combination with othertherapeutic compounds. In this aspect, an amount adequate to accomplishthis, i.e., elicit a response, is defined as “a therapeuticallyeffective combination dose.” The term “simultaneous manner” and“combination treatment” refer to an administration protocol wherein thecompound of the present invention (e.g., compound of Formula I) and atleast one other therapeutic compound (e.g., compound of Formula III, orother therapeutic agent) are administered within a single period oftime. The time period can be measured in hours (e.g., 24 hours) or days(e.g., 30 days).

[0233] “A combination amount sufficient,” “an effective combinationamount” “therapeutically effective combination amount” or “an effectiveamount of the combination of” all refer to a combined amount of both acompound of the present invention (e.g., compound of Formula I) andanother therapeutic compound (e.g., compound of Formula II) or othertherapeutic agent, that is effective to ameliorate symptoms associatedwith FXR-mediated diseases. As used herein, the term “combination” meansthat at least the two compounds can be delivered in a simultaneousmanner, in combination therapy wherein the first compound isadministered first, followed by the second compound, as well as whereinthe second compound is delivered first, followed by the first compound.The desired result can be either a subjective relief of a symptom(s) oran objectively identifiable improvement in the recipient of the dosage.

[0234] In certain preferred aspects, the present invention providescombination therapy for indications such as lipid disorders, skindisorders (e.g. psoriasis), diabetes, cancer, and obesity. For lipiddisorders, the FXR modulators of the present invention are used in acombination dose with statins, including, but not limited to,mevastatin, lovastatin, simvastatin, pravastatin, fluvastatin, andatorvastatin calcium; bile acid sequestrants, including, but not limitedto, cholestryamine and colestipol; probucol; nicotinic acid; andfibrates including, but not limited to, gemfibrozil and clofibrate.

[0235] For skin disorders, the FXR modulators of the present inventionare used in a combination dose with corticosteroids; retinoids (such asetretinate); vitamin D analogs (such as calcipotriene); cyclosporine,and antimetabolites (such as aminopterin and methotrexate).

[0236] For diabetes, the FXR modulators of the present invention areused in a combination dose with insulin; sulfonylureas including, butnot limited to, tolbutamide, acetohexamide, tolazamide, glibenclamide,glyburide, glipizide, and gliclazide; biguanides including, but notlimited to, as metformin; and thiazolidinediones including, but notlimited to, rosiglitazone, troglitazone, and pioglitazone.

[0237] For cancer, the FXR modulators of the present invention are usedin a combination dose with antimitotic agents including, but not limitedto, paclitaxel, vincristine, etoposide, T138067 (Tularik Inc., South SanFrancisco Calif.), and T900607 (Tularik Inc., South San FranciscoCalif.); alkylating agents including, but not limited to,mechlorethamine, cyclophosphamide, and carmustine; antimetabolitesincluding, but not limited to, methotrexate, gemcitabine, lometrexol,5-fluorouracil, and 6-mercaptopurine; cytotoxic antibiotics including,but not limited to, doxorubicin, daunorubicin, bleomycin, mitomycin C,and streptozocin; platinum agents including, but not limited to,cisplatin and carboplatin; hormonal agents including, but not limitedto, anti-estrogens such as tamoxifen and diethylstilbestrol as well asanti-androgens such as flutamide; anti-angiogenesis agents; and farnesyltransferase inhibitors.

[0238] Moreover, for obesity, the FXR modulators of the presentinvention are used in a combination dose with dexfenfluramine,phenylpropanolamine, orlistat, and sibutramine.

EXAMPLES

[0239] Materials and Methods

[0240] Reagents and solvents used below can be obtained from commercialsources such as Aldrich Chemical Co. (Milwaukee, Wis., USA). ¹H-NMRspectra were recorded on a Varian Gemini 400 MHz NMR spectrometer.Significant peaks are tabulated in the order: number of protons,multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m,multiplet; br s, broad singlet) and coupling constant(s) in Hertz (Hz).Electron Ionization (EI) mass spectra were recorded on a Hewlett Packard5989A mass spectrometer. Mass spectrometry results are reported as theratio of mass over charge, followed by the relative abundance of eachion (in parentheses). In tables, a single m/e value is reported for theM+H (or as noted M−H) ion containing the most common atomic isotopes.Isotope patterns correspond to the expected formula in all cases.Electrospray ionization (ESI) mass spectrometry analysis was conductedon a Hewlett-Packard 1100 MSD electrospray mass spectrometer using theHP 1 100 HPLC for sample delivery. Normally the analyte was dissolved inmethanol at 0.1 mg/mL and 1 microliter was infused with the deliverysolvent into the mass spectrometer, which scanned from 100 to 1500daltons. All compounds could be analyzed in the positive ESI mode, using1:1 acetonitrile/water with 1% acetic acid as the delivery solvent. Thecompounds provided below could also be analyzed in the negative ESImode, using 2 mM NH₄OAc in acetonitrile/water as delivery solvent.

Example 1

[0241] This example illustrates the preparation of compound 1.1.4 inFIG. 1A as a representative example of compounds of Formula I.Generally, the compounds of Formula I can be prepared by standard amidecouplings known to those in the art.

[0242] Synthesis of compound 1.1.4:

[0243] 1.1.4

[0244] To a mixture of isophthaloyl dichloride (0.457 g) in methylenechloride was added N-benzyl-p-anisidine (0.85 g), followed by pyridine(330 μL). The reaction was stirred at room temperature and the reactionprogress was monitored by TLC. Upon completion, PS-Trisamine resin (0.74g, loading 4.06 mmol/g, Argonaut) was added and stirred for 1 hour. Thesolution was filtered and the solvent was removed under vacuum. Theresulting solid was recrystallized from hexane and ethyl acetate toprovide the title compound as white crystals (1.25 g, mp 131° C.). ¹HNMR (400 MHz) (DMSO) δ 7.30 (m, 5H); 7.23 (m, 6H); 7.10 (m, 3H), 6.70(m, 8H), 4.99 (s, 4H), 3.60 (s, 6H) MS ESI m/e: 557 (M+H).

Example 2

[0245] This example illustrates the preparation of compound 1.2.23 inFIG. 1C as a representative example of compounds of Formula I. Incertain instances, the compounds of Formula I can be prepared bystandard nucleophilic aromatic substitution reactions (S_(N)Ar) known tothose in the art.

[0246] Synthesis of compound 1.2.23:

[0247] A 10 mL round-bottomed flask was charged with2,3-dichloro-6-nitroquinoxaline (350 mg, 1.47 mmol, prepared accordingto the method of Hinsberg and Pollak: Ber. 1896, 29, 784),3,4-dichloroaniline (710 mg, 4.41 mmol), triethylamine (1.0 mL, 7.35mmol), and 1.0 mL of anhydrous DMF. This mixture was heated to 150° C.for 72 hours. The reaction was then cooled and poured over ice-water.The organics were extracted into EtOAc, dried over MgSO₄, andconcentrated to an orange oil. This oil was purified by flashchromatography (1:1 CH₂Cl₂:hexanes). The desired fractions werecombined, concentrated, and the residue recrystallized from hotEtOAc/hexanes to yield 45 mg of a bright orange solid. An analyticalsample was recrystallized a second time from hot EtOAc/hexanes (m.p. >300° C.). ¹H NMR (400 MHz) (d₆-DMSO) δ 9.66 (s, 1H); 9.53 (s, 1H);8.36 (d, J=1.9 Hz, 1H); 8.29 s, 1H); 8.24 (d, J=1.2 Hz, 1H); 8.17 (dd,J=9.1, 2.2 Hz, 1H); 7.94 (dd, J=9.0, 1.8 Hz, 1H); 7.73-7.66 (m, 3H) MSESI m/e: 492.0 (M−H).

Example 3

[0248] This example illustrates the synthesis of compound 1.2.26:

[0249] Preparation of Compound 1.2.26a.

[0250] A 100 mL round-bottomed flask was charged with2,3-dichloro-5-nitroquinoxaline (7.77g, 31.8 mmol), 3,4-dimethylaniline(7.71 g, 63.6 mmol), triethylamine (22 mL, 159.0 mmol), and anhydrousDMF (20 mL). This was heated at 50° C. overnight. The flask was cooledto room temperature, and the contents were poured over 300 mL ofice-water. This was extracted with 2×200 mL of EtOAc. The organics werecombined and washed with 100 mL 1N HCl, 100 mL H₂O, and 50 mL brine;dried over Na₂SO₄, and concentrated to an orange oily solid in vacuo.The product was further purified using flash chromatography (SiO₂ gel,10% EtOAc/hexanes). The desired fractions were concentrated to yield ayellow solid, which was recrystallized, from hot EtOAc/hexanes to yieldcompound 1.2.30 as yellow crystals (7.12 g, mp 162° C.). ¹H NMR (400MHz) (d₆-DMSO) δ 9.49 (1H, s); 8.60 (1H, d, J=2.6 Hz); 8.35 (1H, dd,J=9.2, 2.6 Hz); 7.77 (1H, d, J=9.2 Hz); 7.59 (2H, m); 7.17 (1H, d, J=8.8Hz); 2.26 (3H, s); 2.23 (3H, s). MS ESI m/e: 327.1 (M−H).

[0251] Conversion of 1.2.26a to 1.2.26.

[0252] A 10 mL pressure tube was charged with compound 1.2.30 (200 mg,0.61 mmol), cyclohexylamine (180 mg, 1.82 mmol), triethylamine (308 mg,3.04 mmol), and anhydrous DMF (0.5 mL). The tube was sealed and heatedat 50° C. for 1.5 hours. The reaction was cooled to room temperature andpoured over 30 mL of ice-water. This was extracted with 2×50 mL ofEtOAc. The combined organics were dried over Na₂SO₄ and concentrated toa red solid. Recrystallization from hot EtOAc gave compound 26 as fineorange crystals (191 mg, mp 223° C.). ¹H NMR (400 MHz) (d₆-DMSO) δ 9.12(1H, s); 8.16 (1H, d, J=2.6 Hz); 7.98 (1H, dd, J=8.9, 2.6 Hz); 7.65 (1H,dd, J=8.1, 2.2 Hz); 7.57 (1H, d, J=1.8 Hz); 7.53 (1H, d, J=8.9 Hz); 7.36(1H, d, J=6.9 Hz); 7.17 (1H, d, J=8.2 Hz); 4.20-4.09 (1H, m); 2.27 (3H,s); 2.23 (3H, s); 2.11-2.08 (2H, m); 1.83-1.79 (2H, m); 1.70-1.66 (1H,m); 1.45-1.25 (5H, m). MS ESI m/e: 398.1 (M−H).

Example 4

[0253] This example illustrates the preparation of compound 2.1.1 inFIG. 2A as a representative example of compounds of Formula II.Generally, the compounds of Formula II can be prepared by standardmethods known to those in the art.

[0254] Synthesis of compound 2.1.1:

[0255] N-(2,6-dihydoxy-phenyl)-benzamide (7 g, 30.5 mmol, described inChem. Ber. 1958, 91, 1123-1125) was suspended in 85% H₃PO₄ (150 mL) and2-methyl-2-propanol (95 mL, 68.4 g, 922 mmol). The reaction mixture wasvigorously stirred for 16 hours and then poured into 500 mL of deionizedwater and extracted with 500 mL of EtOAc. The aqueous layer wascarefully treated with 50 g of NaHCO₃ to partially neutralize thephosphoric acid and aid in layer separation. The aqueous layer was againextracted with 200 mL of EtOAc. The combined organic layers were washed1×100 mL sat. brine, dried over Na₂SO₄, filtered, and concentrated to atan amorphous solid. Trituration with CH₂Cl₂/hexanes resulted in theformation of a fine white solid, which was collected by filtration,rinsed with hexanes, and dried under vacuum. The product (8.9 g, 85%yield, mp 197° C.) was found to be >95% pure by HPLC. ¹H NMR (400 MHz)(CD₃CN) δ 8.49 (bs, 1H); 8.05 (d, J=7.2 Hz, 2H); 7.64 (t, J=6.5 Hz, 1H);7.57 (t, J=7.0 Hz, 2H); 7.18 (s, 1H); 6.91 (s, 2H); 1.39 (s, 18H). MSESI m/e: 342.2 (M+H), 364.1 (M+Na)

Example 5

[0256] This example illustrates the synthesis of compound 2.2.10 in FIG.2B:

[0257] N-(3-trifluoromethyl-phenyl)-benzenesulfonamide (2 g, 6.6 mmol,described in Coll. Chech. Chem. Comm. 1987, 52, 2900-2908.) wasdissolved in anhydrous THF (6 mL) and then treated with potassiumtert-butoxide (6.5 mL of 1.0 N solution in THF). After five min.,chloroacetamide 2.2.10a (1.4 g, 6.2 mmol, described in J. Ind. Chem.Soc. 1963, 40, 885-888.) was added in one portion and Bu₄NI (229 mg,0.62 mmol) was added to catalyze the alkylation. The reaction mixturewas warmed to 50° C. After 16 h, the reaction was complete by TLCanalysis. The reaction mixture was poured into sat. NH₄Cl_((aq)) anddiluted with 1:1 ethyl acetate/hexanes. The aqueous phase was extracted1×50 mL ethyl acetate and the combined organic layers were washed withsat. brine, dried over Na₂SO₄, filtered, and concentrated to a lightyellow oil. Purification by flash chromatography (silica gel, elutingwith 40% ethyl acetate/hexanes) afforded an amorphous solid, which couldbe crystallized only from ether/hexanes at −20° C. After filtration,rinsing with hexanes, and drying under vacuum, compound 2.2.10 wasobtained as a fine white powder (2.1 g, m. p. 93° C.). ¹H NMR (400 MHz)(CDCl₃) δ 7.64 (t, J=7.6 Hz, 1H); 7.62-7.46 (m, 5H); 7.41 (t, J=8.0 Hz,1H); 7.29 (s, 1H); 7.20 (d, J=8.0 Hz, 1H); 7.10 (d, J=8.8 Hz, 2H); 6.82(d, J=8.4 Hz, 2H); 6.60 (d, J=8.4 Hz, 1H); 5.00 (quintet, J=7.2 Hz, 1H);4.28 (d, J=16.8 Hz, 1H); 4.12 (d, J=16.8 Hz, 1H); 3.79 (s, 3H); 1.44 (d,J=6.8 Hz, 3H). MS ESI m/e: 493.1 (M+H), 515.2 (M+Na).

Example 6

[0258] This example illustrates the preparation of compound 3.1.1 inFIG. 3A as a representative example of compounds of Formula III.Generally, the compounds of Formula III can be prepared by standardamide couplings known to those in the art.

[0259] Synthesis of compound 3.1.1:

[0260] N-phthaloyl-DL-phenylalanine (8.23 g, 0.028 mol, described in J.Am. Chem. Soc. 1948, 70, 1473) was treated with POCl₃ (25 ml, 0.27 mol)and the resultant mixture heated at 75° C. for 3 hours, during whichtime the reaction mixture became a yellow solution. Excess POCl₃ wasremoved (by rotary evaporation followed by high vacuum pump for 1 hour)and the residue dissolved in dichloromethane (100 ml) and treated withdicyclohexylamine (8.5 ml, 0.04 mol). The reaction mixture was stirredat room temperature for 4 hours before being filtered; the solvent wasevaporated and the crude product purified by flash column chromatography(eluant: 50% ethyl acetate in hexanes) to afford the title compound as awhite solid (7.08 g, mp 183° C.).

[0261]¹H NMR (400 MHz) (CDCl₃): δ 7.78 (m, 2H); 7.70 (m, 2H); 7.19 (m,5H); 5.26 (m, 1H); 3.65 (m, 1H); 3.41 (m, 1H); 2.88 (m, 1H); 2.57 (m,2H); 1.34 (m, 19H) ppm. MS ESI m/e: 459.2 (M+H).

Example 7

[0262] This example illustrates the preparation of compound 3.2.3 inFIG. 3C as a representative example of compounds of Formula III.Generally, the compounds of Formula IV can be prepared by standard amidecouplings known to those in the art.

[0263] Conversion of 3.1.1 to 3.2.3a.

[0264] To a solution of the phthalimide substrate A (5.70 g, 0.012 mol)in 2-propanol (75 ml) and water (10.5 ml) was added NaBH₄ (2.33 g, 0.06mol) and the reaction mixture stirred at room temperature overnight.Acetic acid (12.5 ml) was added dropwise, the reaction flask wasstoppered and the mixture heated to 80° C. for 4 hours. The reactionmixture was cooled to room temperature, poured into water andneutralised with solid sodium bicarbonate. The aqueous mixture wasextracted with ethyl acetate, the combined organic phase dried and thesolvent evaporated. The product was used without further purification.MS ESI m/e: 329.3 (M+H).

[0265] Conversion of 3.2.3a to 3.2.3b.

[0266] To a solution of crude 3.2.3a (203 mg, 0.62 mmol) indichloromethane (5 ml) at 0° C. was added triethylamine (0.1 ml, 0.72mmol) followed by benzoyl chloride (0.085 ml, 0.73 mmol) and thereaction mixture allowed to warm to room temperature and stirred for 3hours. The reaction was quenched by the addition of 0.05 M NaOH (aq) andthe aqueous layer extracted with dichloromethane. The combined organicphase was dried (Na₂SO₄), the solvent evaporated and the crude productpurified by flash column chromatography (eluant: 20% ethyl acetate inhexanes). MS ESI m/e: 433.3 (M+H), 455.3 (M+Na).

[0267] Conversion of 3.2.3b to 3.2.3.

[0268] To 3.2.3b (62 mg, 0.14 mmol) in a 1:1 mixture of THF and DMF (20ml) at 0° C. was added sodium hydride (8 mg, 0.20 mmol of a 60%dispersion in mineral oil) and the resultant mixture stirred at 0° C.for 20 minutes before being allowed to warm to room temperature.Iodomethane (0.014 ml, 0.22 mmol) was then added to the reaction mixtureand the resultant mixture stirred at room temperature for 4 h. Water wasadded to quench the reaction and the aqueous layer was extracted withethyl acetate. The combined organic phase was dried (MgSO₄), the solventevaporated in vacuo and the crude product purified by flash columnchromatography (eluant: 20% ethyl acetate in hexanes) to afford compound3.2.3 as a white solid (58 mg, mp 123.1° C.). ¹H NMR (400 MHz) (CDCl₃):δ 7.29 (m, 8H); 6.99 (m, 2H); 5.94 (t, J=8.0 Hz, 1H); 3.89 (m, 1H); 3.33(m, 1H); 3.12 (m, 1H); 2.95 (m, 1H); 2.85 (s, 3H); 2.52 (m, 2H); 1.42(m, 18H). MS ESI m/e: 447.4 (M+H).

Example 8

[0269] This example illustrates the preparation of compound 4.7 in FIG.4A as a representative example of compounds of Formula IV. Generally,the compounds of Formula V can be prepared by standard amide couplingsknown to those in the art.

[0270] Synthesis of compound 4.7:

[0271] To a solution of N-phthaloyl-DL-valine (306 mg, 1.24 mmol,described in J. Am. Chem. Soc. 1948, 70, 1473) in DMF (10 mL) was added1-hydroxy-7-azabenzotriazole (166 mg, 1.22 mmol) and 1,6-hexanediamine(76 mg, 0.65 mmol), followed by O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (461 mg, 1.21 mmol) andN-methylmorpholine (0.27 mL, 2.46 mmol) and the reaction mixture stirredat room temperature for 3 hours. The reaction mixture was diluted withCH₂Cl₂ (10 mL) and added to water (100 mL); the aqueous layer wasextracted with CH₂Cl₂ and the combined organic phase evaporated. Thecrude product was dried (MgSO₄) and purified by flash columnchromatography (eluant: 50% ethyl acetate in hexanes) to afford thetitle compound as a white solid (84 mg, mp 165s° C.). ¹H NMR (400 MHz)(CDCl₃): δ 7.88 (m, 4H); 7.73 (m, 4H); 7.04 (br, s, 2H); 4.39 (m, 2H);3.28 (m, 4H); 2.79 (m, 2H); 1.38 (m, 8H); 1.09 (m, 6H); 0.85 (m, 6H)ppm. MS ESI m/e: 547.2 (M+H).

Example 9

[0272] This example illustrates the preparation of compound 5.3 in FIG.5 as a representative example of compounds of Formula V. Generally, thecompounds of Formula V can be prepared by standard methods known tothose in the art.

[0273] Preparation of sulfonamide 5.3a. 2,5-dimethyl-aniline (2.4 g, 20mmol) and 2-nitrobenzenesulfonyl chloride (4.4 g, 20 mmol) are dissolvedin CH₂Cl₂ and treated with pyridine (1.8 mL, 22 mmol). The reactionmixture is stirred at room temperature until TLC shows no startingmaterial. The reaction mixture is quenched with dilute HCl, and theorganic layer washed with saturated brine. Drying over MgSO₄,filtration, and concentration provides a solid that is recrystallizedfrom hot EtOAc/hexanes to provide2-nitro-N-(2,5-dimethyl-phenyl)-benzenesulfonamide.

[0274] Conversion of 5.3a to 5.3b.

[0275] 2-nitro-N-(2,5-dimethyl-phenyl)-benzenesulfonamide (6.1 g, 20mmol) in EtOAc (60 mL) is treated with SnCl₂-2H₂O (22.5 g, 100 mmol).The reaction mixture is heated to reflux for 30 minutes and then allowedto cool to room temperature. 2 N KOH (250 mL, 500 mmol) is added and theresulting mixture stirred vigorously until all solids dissolve. Theaqueous layer is separated and extracted 2×100 mL EtOAc. The combinedorganic layers are washed with sat. brine, dried over MgSO₄, filtered,and concentrated to a solid, which is recrystallized from hotEtOAc/hexanes to provide2-amino-N-(2,5-dimethyl-phenyl)-benzenesulfonamide.

[0276] Conversion of 5.3b to 5.3c.

[0277] Conversion of 2-amino-N-(2,5-dimethyl-phenyl)-benzenesulfonamideto 1,4-Dimethyl-10H-9-thia-10-aza-phenanthrene 9,9-dioxide is carriedout by diazotization according to the method of Ullmann and Gross (Ber.Dtsch. Chem. Ges. 1910, 43, 2694).

[0278] Conversion of 5.3c to 5.3.

[0279] 1,4-Dimethyl-10H—9-thia-10-aza-phenanthrene 9,9-dioxide (2.6 g,10 mmol) is dissolved in pyridine (10 mL) and treated withp-toluenesulfonyl chloride (2.8 g, 15 mmol). The reaction mixturestirred at 60° C. until TLC shows disappearance of the startingmaterial. The reaction mixture is allowed to cool and then poured into100 mL of 3N HCl. The mixture is extracted 2×100 mL EtOAc and thecombined organic layers are washed with sat. brine, dried over MgSO₄,filtered, and concentrated to a solid, which is recrystallized from hotEtOAc/hexanes to give1,4-dimethyl-10-(toluene-4-sulfonyl)-10H-9-thia-10-aza-phenanthrene9,9-dioxide.

Example 10

[0280] Compounds of the present invention (illustrated by the examplesabove and in the figures) were evaluated for FXR binding activity. IC₅₀values were measured, representing the concentration of the compound atwhich 50% of the activity is inhibited. A substantial number of thecompounds exhibited IC₅₀ values ranging from less than 1 to about 30 μMor more. In preferred embodiments of the invention, the compoundsexhibited IC₅₀ values of 10 μM or more. In more preferred embodiments,the compounds exhibited IC₅₀ values ranging from 1 μM to about 10 μM. Inthe most preferred embodiments, the compounds exhibited IC₅₀ values ofless than 1 μM.

Example 11

[0281] This Example describes an in vitro assay that is useful toidentify compounds that modulate binding of FXR ligands to the FXRligand binding domain. The ability of compounds of Formulae I-V toaffect the binding of a labeled sensor peptide that is derived from thecoactivator SRC-1 to an FXR ligand binding domain was tested.

[0282] Fluorescence polarization was used to study the effect ofcompounds of Formulae I-V on the ability of the FXR LBD to bind a sensorpeptide. The assay reagents were as follows:

[0283] Sensor: Rhodamine-labeled ILRKLLQE peptide (final conc.=1-5 nM).It is noted that the Rhodamine-labeled peptide comprises, at a minimum,the following sequence LXXLLXX, wherein X is any amino acid. Additionalamino acids can be added to both the N-terminus and the C-terminus ofthis core peptide. In preferred embodiments, the peptide is 8 aminoacids in length and, more preferable, about 11 amino acids in length.

[0284] Receptor: Glutathione-S-transferase/FXR ligand binding domainfusion protein (final conc.=100-200 nM).

[0285] Buffer: 10 mM HEPES, 10 mM NaCl, 6 mM magnesium chloride, pH 7.6.

[0286] Protocol

[0287] 1. Add 90 microliters of Sensor/Receptor mixture to each well ofa 96-well microtiter plate.

[0288] 2. Add 10 microliters of test compound per well.

[0289] 3. Shake 5 minutes and within 5 minutes determine the amount offluorescence polarization by using a Fluorolite FPM-2 FluorescencePolarization Microtiter System (Dynatech Laboratories, Inc.)

[0290] Ten ng/μL of GST-FXR fusion protein was mixed with arhodamine-labeled peptide comprising LXXLLXX, wherein X is any aminoacid, and the panel of bile acids (concentrations as indicated).Fluorescence polarization was read after a room temperature incubationand brief shaking. Change in millipolarization (mP) units is thedifference treated and untreated samples. The high change in mP unitsdemonstrates that the labeled peptide binds to GST-FXR in aCDCA-dependent manner.

[0291] Using fluorescence resonance energy transfer (FRET) one can studythe effect of different compounds of Formula I-V on the ability of FXRto bind the coactivator SRC-1.

[0292] All publications and patent applications cited in thisspecification are herein incorporated by reference as if each individualpublication or patent application were specifically and individuallyindicated to be incorporated by reference. Although the foregoinginvention has been described in some detail by way of illustration andexample for purposes of clarity of understanding, it will be readilyapparent to those of ordinary skill in the art in light of the teachingsof this invention that certain changes and modifications may be madethereto without departing from the spirit or scope of the appendedclaims.

What is claimed is:
 1. A compound of the formula B¹-L¹-A¹-L²-B²  Iwherein: A¹ is a member selected from the group consisting of alkylene,alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene,heteroarylene, heterocycloalkylene, and heterocycloalkenylene, or,alternatively, A¹ represents a single or double bond linking L¹ and L²;L¹ and L² are each independently a member selected from the groupconsisting of O—, —S—, —N(R¹)-, —C(O)—, —C(O)N(R¹)-, —O-alkylene-,—S-alkylene-, —N(R¹)-alkylene, —C(O)-alkylene, —C(O)N(R)-alkylene,—C(O)—O-alkylene, alkylene, alkenylene, alkynylene, cycloalkylene,cycloalkenylene, arylene, heteroarylene, heterocycloalkylene, andheterocycloalkenylene; B¹ and B² are each independently a memberselected from the group consisting of alkyl, cycloalkyl, cycloalkenyl,aryl, heteroaryl, heterocycloalkyl, and heterocycloalkenyl;alternatively, L¹ can be additionally linked to B¹ via a group X¹ toform a 5-9 member ring; and L² can be additionally linked to B² via agroup X² to form a 5-9 member ring; X¹ and X² are each independently amember selected from the group consisting of —O—, —S—, —N(R²)—, —C(O)—,—C(O)N(R²)—, —O-alkylene, —S-alkylene, —N(R²)-alkylene, —C(O)-alkylene,—C(O)N(R²)-alkylene, and —C(O)—O-alkylene; and R¹ and R² are eachindependently a member selected from the group consisting of hydrogen,alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, aryl(heteroalkyl),(heteroaryl)alkyl, and (heteroaryl)heteroalkyl.
 2. The compound of claim1, wherein A¹ is a member selected from the group consisting of(C₁-C₈)alkylene, arylene, heteroarylene and a single bond; L¹ and L² areeach independently a member selected from the group consisting of —C(O)—and —C(O)N(R¹)-; R¹ is a member selected from the group consisting of(C₅-C₈)cycloalkyl, aryl, heteroaryl, aryl(C₁-C₄)alkyl, and(heteroaryl)(C₁-C₄)alkyl; and B¹ and B² are each independently a memberselected from the group consisting of aryl, heteroaryl,aryl(C₁-C₄)alkyl, (heteroaryl)(C₁-C₄)alkyl, (C₁-C₈)alkyl, and(C₅-C₈)cycloalkyl.
 3. The compound of claim 1, wherein A¹ is a memberselected from the group consisting of (C₁-C₈)alkylene, phenylene,divalent pyridine and a single bond; L¹ and L² are each independently amember selected from the group consisting of —C(O)— and —C(O)N(R¹)-; R¹is optionally substituted (C₅-C₈)cycloalkyl, optionally substitutedphenyl, optionally substituted benzyl, and (C₁-C₈)alkyl; and B¹ and B²are each independently a member selected from the group consisting ofoptionally substituted (C₅-C₈)cycloalkyl, optionally substituted phenyl,and optionally substituted benzyl.
 4. The compound of claim 1, whereinA¹ is a member selected from the group consisting of alkylene, arylene,heteroarylene and a single bond; L¹ and L² are each —C(O)N(R¹)—; R¹ is amember selected from the group consisting of aryl, heteroaryl,arylalkyl, and (heteroaryl)alkyl; and B¹ and B² are each independently amember selected from the group consisting of aryl, heteroaryl,arylalkyl, (heteroaryl)alkyl, alkyl, and cycloalkyl.
 5. The compound ofclaim 1, wherein A¹ is a heteroarylene group containing two fused rings;L¹ and L² are each independently a member selected from the groupconsisting of —O—, —NH—, and —N(R¹)-; R¹ is a member selected from thegroup consisting of alkyl and heteroalkyl; and B¹ and B² are eachindependently a member selected from the group consisting of aryl,heteroaryl, arylalkyl, (heteroaryl)alkyl, alkyl, and cycloalkyl.
 6. Acompound of the formula

wherein: A² and A³ are each independently a member selected from thegroup consisting of alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, arylalkyl,(heteroaryl)alkyl, aryl(heteroalkyl), and (heteroaryl)heteroalkyl; B³ isa member selected from the group consisting of hydrogen,-alkylene-C(O)R³, —C(O)R³, alkyklene-C(O)N(R³R⁴), —C(O)N(R³R⁴),alkylene-S(O)_(n)N(R³R⁴), -S(O)_(n)N(R³R⁴), alkylene-N(R³R⁴),alkylene-OR³, and —C(O)OR³; R³ and R⁴ are each independently a memberselected from the group consisting of hydrogen, alkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl,heteroaryl, arylalkyl, (heteroaryl)alkyl, aryl(heteroalkyl), and(heteroaryl)heteroalkyl; X is a member selected from the groupconsisting of C, S, and N; and the subscripts n and p are eachindependently an integer from 0-2, provided that the following compoundis excluded:


7. The compound of claim 6, wherein A² and A³ are each independently amember selected from the group consisting of aryl and heteroaryl; B³ isa member selected from the group consisting of alkylene-C(O)N(R³R⁴), andalkylene-S(O)_(n)N(R³R⁴); wherein R³ is arylalkyl or (heteroaryl)alkyl;R⁴ is hydrogen; X is S; and n is
 2. 8. The compound of claim 6, whereinA² is an aryl group substituted ortho to the nitrogen with a memberselected from the group consisting of —OH, —NH₂, —NHC(O)-alkyl,—NHSO₂-alkyl; A³ is a member selected from the group consisting of aryland heteroaryl; B³ is hydrogen; X is C; and p is
 1. 9. A compound of theformula:

wherein: A⁴ is a member selected from the group consisting of hydrogen,—C(O)R⁵, —C(O)N(R⁵R⁶), —S(O)_(n)N(R⁵R⁶), -alkylene-N(R⁵R⁶),-alkylene-OR⁵ and —C(O)OR⁵; L³ and L⁴ are each independently a memberselected from the group consisting of a single bond, —C(O)—, —S(O)_(p)—,and alkylene, wherein the subscript p is an integer from 0-2; B⁴, B⁵ andB⁶ are each independently a member selected from the group consisting ofhydrogen, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,fused-benzoheterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl,heteroaryl, arylalkyl, aryl(heteroalkyl), (heteroaryl)alkyl, and(heteroaryl)heteroalkyl; alternatively, B⁴ and B⁵ join to form adivalent arylene, heteroarylene, alkylene, or cycloalkylene linkagebetween L³ and L⁴, and B⁶ is a member selected from the group consistingof hydrogen, alkyl, heteroalkyl, heterocycloalkyl, arylalkyl, or(heteroaryl)alkyl. X³ and Y are each independently a trivalent nitrogenatom or a trivalent or tetravalent carbon atom; and R⁵ and R⁶ are eachindependently a member selected from the group consisting of hydrogen,alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, aryl(heteroalkyl),(heteroaryl)alkyl, and (heteroaryl)heteroalkyl.
 10. The compound ofclaim 9, wherein A⁴ is a member selected from the group consisting ofhydrogen, —C(O)N(R⁵R⁶) and —S(O)₂N(R⁵R⁶); R⁵ and R⁶ are eachindependently a member selected from the group consisting of alkyl,cycloalkyl, and heterocycloalkyl; L³ and L⁴ are each independently amember selected from the group consisting of —C(O)—, —S(O)₂—, and loweralkylene; B⁴ and B⁵ join to from an arylene or heteroarylene linkagebetween L³ and L⁴; X is tetravalent carbon in the R configuration; Y istrivalent nitrogen; and B⁶ is a member selected from the groupconsisting of hydrogen, alkyl, heteroalkyl, heterocycloalkyl, arylalkyl,or (heteroaryl)alkyl.
 11. The compound of claim 9, wherein A⁴ is amember selected from the group consisting of hydrogen, —C(O)N(R⁵R⁶) and—S(O)₂N(R⁵R⁶); R⁵ and R⁶ are each independently a member selected fromthe group consisting of alkyl, cycloalkyl, and heterocycloalkyl; L³ andL⁴ are each independently a member selected from the group consisting of—C(O)—, —S(O)²-, and lower alkylene; B⁴ and B⁵ are each independently amember selected from the group consisting of hydrogen, alkyl, arylalkyl,aryl, and heteroaryl; X is tetravalent carbon in the R configuration; Yis trivalent nitrogen; and B⁶ is a member selected from the groupconsisting of hydrogen, alkyl, heteroalkyl, heterocycloalkyl, arylalkyl,and (heteroaryl)alkyl.
 12. The compound of claim 9, said compound havingthe formula

wherein: X⁴, X⁵ and X⁶ are each independently C or S; R¹⁰ and R¹¹ areeach independently alkyl, cycloalkyl, or heterocycloalkyl; R⁹ is anoptionally substituted aryl, heteroaryl, arylalkyl, (heteroaryl)alkyl,heterocycloalkyl; R¹⁴ is selected from hydrogen, halogen, alkyl, alkoxy,alkylamino, alkylthio, acyl, cycloalkyl and aryl; and the subscripts p,q, and r are each independently integers from 0-2.
 13. A compound of theformula:

wherein: A⁴ is a member selected from the group consisting of hydrogen,—C(O)R⁵, —C(O)N(R⁵R⁶), —S(O)_(n)N(R⁵R 6), -alkylene-N(R⁵R⁶),-alkylene-OR⁵ and —C(O)OR⁵; B⁵ and B⁶ are members independently selectedfrom the group consisting of hydrogen, alkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, fused-benzoheterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, aryl(heteroalkyl),(heteroaryl)alkyl and (heteroaryl)heteroalkyl; and X³ is a trivalentnitrogen atom or a trivalent or tetravalent carbon atom.
 14. A compoundof the formula:

wherein: A⁵ is a member selected from the group consisting of —C(O)—,-alkylene-, —S(O)_(n)—, —C(O)N(R¹²)-, —S(O)₂N(R 2)-, -alkylene-N(R¹²)-,-alkylene-O—, and —C(O)O—; L⁵ and L⁶ are each independently a memberselected from the group consisting of —C(O)—, —S(O)_(n)—; and alkylene,wherein the subscript n is an integer from 0-2; B⁷, B⁸, and B⁹ are eachindependently a member selected from the group consisting of alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, benzoheterocycloalkyl,cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, arylalkyl,aryl(heteroalkyl), (heteroaryl)alkyl, and (heteroaryl)heteroalkyl;alternatively, B⁷ and B⁸ join to form a divalent arylene, heteroarylene,alkylene, or cycloalkylene linkage between L⁵ and L⁶; Z is a memberselected from the group consisting of alkylene, heteroalkylene,cycloalkylene, and heterocycloalkylene; X⁷ and Y¹ are each independentlya trivalent nitrogen atom or a trivalent or tetravalent carbon atom; andR¹² is a member selected from the group consisting of hydrogen, alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, aryl(heteroalkyl),(heteroaryl)alkyl, and (heteroaryl)heteroalkyl.
 15. The compound ofclaim 14, wherein A⁵ is a member selected from the group consistingof-C(O)—, —C(O)N(R¹²)- and —S(O)₂N(R¹²)-; R¹² is a member selected fromthe group consisting of alkyl, cycloalkyl, and heterocycloalkyl; B⁷ andB⁸ are joined in an arylene or heteroarylene linkage between L⁵ and L⁶;B⁹ is a member selected from the group consisting of alkyl, heteroalkyl,heterocycloalkyl, arylalkyl, and (heteroaryl)alkyl; Z is alkylene,heteroalkylene, or heterocycloalkylene; L⁵ and L⁶ are each independentlya member selected from the group consisting of-C(O)—, —S(O)₂—, or loweralkylene; X⁷ is tetravalent carbon; and Y¹ is trivalent nitrogen.
 16. Acompound of the formula:

wherein: A⁶ and A⁷ are each independently a member selected from thegroup consisting of arylene, heteroarylene, cycloalkylene, andheterocycloalkylene; B¹⁰ is a member selected from the group consistingof aryl, heteroaryl, arylalkyl, (heteroaryl)alkyl, alkyl, cycloalkyl,cycloalkenyl, heteroalkyl, heterocycloalkyl, and heterocycloalkenyl; L⁷,L⁸, and L⁹ are each independently a member selected from the groupconsisting of —O—, —S—, —N(R¹³), —C(O)—, —S(O)—, —S(O)₂—, alkylene,—O-alkylene, —S-alkylene, —N(R¹³)-alkylene, —C(O)-alkylene,—C(O)N(R¹³)-alkylene, —C(O)—O-alkylene, a single bond, and a doublebond; X⁸ is a member selected from the group consisting of N, and CR¹³;and R¹³ is a member selected from the group consisting of hydrogen,alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, arylalkyl, and (heteroaryl)alkyl.17. The compound of claim 16, wherein A⁶ and A⁷ are each independently amember selected from the group consisting of aryl, heteroaryl,cycloalkyl, and heterocycloalkyl; B¹⁰ is a member selected from thegroup consisting of aryl, heteroaryl, arylalkyl, and (heteroaryl)alkyl;L⁷ and L⁸ are each independently a member selected from the groupconsisting of —C(O)—, —S(O)—, and —S(O)₂-; L⁹ is a member selected fromthe group consisting of —C(O)—, alkylene, and a single bond; and X⁵ isN.
 18. A pharmaceutical composition, said pharmaceutical compositioncomprising: a) a compound of claim 1; and b) a pharmaceuticallyacceptable carrier or excipient.
 19. A pharmaceutical composition, saidpharmaceutical composition comprising: a) a compound of the formula

wherein: A² and A³ are each independently a member selected from thegroup consisting of alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, arylalkyl,(heteroaryl)alkyl, aryl(heteroalkyl), and (heteroaryl)heteroalkyl; B³ isa member selected from the group consisting of hydrogen,-alkylene-C(O)R³, —C(O)R³, alkyklene-C(O)N(R³R⁴), —C(O)N(R³R⁴),alkylene-S(O)_(n)N(R³R⁴), —S(O)_(n)N(R³R⁴), alkylene-N(R³R⁴),alkylene-OR³, and —C(O)OR³; R³ and R⁴ are each independently a memberselected from the group consisting of hydrogen, alkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl,heteroaryl, arylalkyl, (heteroaryl)alkyl, aryl(heteroalkyl), and(heteroaryl)heteroalkyl; X is a member selected from the groupconsisting of C, S, and N; and the subscripts n and p are eachindependently an integer from 0-2; and b) a pharmaceutically acceptablecarrier or excipient.
 20. A pharmaceutical composition, saidpharmaceutical composition comprising: a) a compound of claim 9; and b)a pharmaceutically acceptable carrier or excipient.
 21. A pharmaceuticalcomposition, said pharmaceutical composition comprising: a) a compoundof claim 13; and b) a pharmaceutically acceptable carrier or excipient.22. A pharmaceutical composition, said pharmaceutical compositioncomprising: a) a compound of claim 14; and b) a pharmaceuticallyacceptable carrier or excipient.
 23. A pharmaceutical composition, saidpharmaceutical composition comprising: a) a compound of claim 16; and b)a pharmaceutically acceptable carrier or excipient.
 24. A method fortreating a FXR-mediated disease in a mammal, said method comprising:administering a compound of claim 1, thereby treating a FXR-mediateddisease in a mammal.
 25. A method for treating a FXR-mediated disease ina mammal, said method comprising: administering a compound of theformula

wherein: A² and A³ are each independently a member selected from thegroup consisting of alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, arylalkyl,(heteroaryl)alkyl, aryl(heteroalkyl), and (heteroaryl)heteroalkyl; B³ isa member selected from the group consisting of hydrogen,-alkylene—C(O)R³, —C(O)R³, alkyklene-C(O)N(R³R⁴), —C(O)N(R³R⁴),alkylene-S(O)_(n)N(R³R⁴), —S(O)_(n)N(R³R⁴), alkylene-N(R³R⁴),alkylene-OR³, and —C(O)OR³; R³ and R⁴ are each independently a memberselected from the group consisting of hydrogen, alkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl,heteroaryl, arylalkyl, (heteroaryl)alkyl, aryl(heteroalkyl), and(heteroaryl)heteroalkyl; X is a member selected from the groupconsisting of C, S, and N; and the subscripts n and p are eachindependently an integer from 0-2; thereby treating a FXR-mediateddisease in a mammal.
 26. A method for treating a FXR-mediated disease ina mammal, said method comprising: administering a compound of claim 9,thereby treating a FXR-mediated disease in a mammal.
 27. A method fortreating a FXR-mediated disease in a mammal, said method comprising:administering a compound of claim 13, thereby treating a FXR-mediateddisease in a mammal.
 28. A method for treating a FXR-mediated disease ina mammal, said method comprising: administering a compound of claim 14,thereby treating a FXR-mediated disease in a mammal.
 29. A method fortreating a FXR-mediated disease in a mammal, said method comprising:administering a compound of claim 16, thereby treating a FXR-mediateddisease in a mammal.
 30. A method for modulating cyp7a expression levelsin a mammal, said method comprising: administering a compound of claim1, thereby modulating cyp7a expression levels in a mammal.
 31. A methodfor modulating cyp7a expression levels in a mammal, said methodcomprising: administering a compound of the formula

wherein: A² and A³ are each independently a member selected from thegroup consisting of alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, arylalkyl,(heteroaryl)alkyl, aryl(heteroalkyl), and (heteroaryl)heteroalkyl; B³ isa member selected from the group consisting of hydrogen,-alkylene-C(O)R³, —C(O)R³, alkyklene-C(O)N(R³R⁴), —C(O)N(R³R⁴),alkylene-S(O)_(n)N(R³R⁴), —S(O)_(n)N(R³R⁴), alkylene-N(R³R⁴),alkylene-OR³, and —C(O)OR³; R³ and R⁴ are each independently a memberselected from the group consisting of hydrogen, alkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl,heteroaryl, arylalkyl, (heteroaryl)alkyl, aryl(heteroalkyl), and(heteroaryl)heteroalkyl; X is a member selected from the groupconsisting of C, S, and N; and the subscripts n and p are eachindependently an integer from 0-2; thereby modulating cyp7a expressionlevels in a mammal.
 32. A method for modulating cyp7a expression levelsin a mammal, said method comprising: administering a compound of claim9, thereby modulating cyp7a expression levels in a mammal.
 33. A methodfor modulating cyp7a expression levels in a mammal, said methodcomprising: administering a compound of claim 13, thereby modulatingcyp7a expression levels in a mammal.
 34. A method for modulating cyp7aexpression levels in a mammal, said method comprising: administering acompound of claim 14, thereby modulating cyp7a expression levels in amammal.
 35. A method for modulating cyp7a expression levels in a mammal,said method comprising: administering a compound of claim 16, therebymodulating cyp7a expression levels in a mammal.